Amido ether substituted imidazoquinolines

ABSTRACT

Imidazoquinoline and tetrahydroimidazoquinoline compounds that contain ether and amide functionality at the 1-position are useful as immune response modifiers. The compounds and compositions of the invention can induce the biosynthesis of various cytokines and are useful in the treatment of a variety of conditions including viral diseases and neoplastic diseases.

This application claims the benefit of previously filed ProvisionalApplication Serial No. 60/254,218, filed on Dec. 8, 2000.

FIELD OF THE INVENTION

This invention relates to imidazoquinoline compounds that have ether andamido functionality at the 1-position, and to pharmaceuticalcompositions containing such compounds. A further aspect of thisinvention relates to the use of these compounds as immunomodulators, forinducing cytokine biosynthesis in animals, and in the treatment ofdiseases, including viral and neoplastic diseases.

BACKGROUND OF THE INVENTION

The first reliable report on the 1H-imidazo[4,5-c]quinoline ring system,Backman et al., J. Org. Chem. 15, 1278-1284 (1950) describes thesynthesis of1-(6-methoxy-8-quinolinyl)-2-methyl-1H-imidazo[4,5-c]quinoline forpossible use as an antimalarial agent. Subsequently, syntheses ofvarious substituted 1H-imidazo[4,5-c]quinolines were reported. Forexample, Jain et al., J. Med. Chem. 11, pp. 87-92 (1968), synthesizedthe compound 1-[2-(4-piperidyl)ethyl]-1H-imidazo[4,5-c]quinoline as apossible anticonvulsant and cardiovascular agent. Also, Baranov et al.,Chem. Abs. 85, 94362 (1976), have reported several2-oxoimidazo[4,5-c]quinolines, and Berenyi et al., J. Heterocyclic Chem.18, 1537-1540 (1981), have reported certain2-oxoimidazo[4,5-c]quinolines.

Certain 1H-imidazo[4,5-c]quinolin-4-amines and 1- and 2-substitutedderivatives thereof were later found to be useful as antiviral agents,bronchodilators and immunomodulators. These are described in, interalia, U.S. Pat. Nos. 4,689,338; 4,698,348; 4,929,624; 5,037,986;5,268,376; 5,346,905; and 5,389,640, all of which are incorporatedherein by reference.

There continues to be interest in the imidazoquinoline ring system.

Certain 1H-imidazo[4,5-c]naphthyridine-4-amines,1H-imidazo[4,5-c]pyridin-4-amines, and1H-imidazo[4,5-c]quinolin-4-amines having an ether containingsubstituent at the 1 position are known. These are described in U.S.Pat. Nos. 5,268,376; 5,389,640; 5,494,916; and WO 99/29693.

Despite these attempts to identify compounds that are useful as immuneresponse modifiers, there is a continuing need for compounds that havethe ability to modulate the immune response, by induction of cytokinebiosynthesis or other mechanisms.

SUMMARY OF THE INVENTION

We have found a new class of compounds that are useful in inducingcytokine biosynthesis in animals. Accordingly, this invention providesimidazo[4,5-c]quinoline-4-amine and tetrahydroimidazo[4,5-c]quinoline-4-amine compounds that have an ether containingsubstituent at the 1-position. The compounds are defined by Formulas (I)and (II), which are defined in more detail infra. These compounds sharethe general structural formula:

wherein X, R₁, R₂, and R are as defined herein for each class ofcompounds having Formulas (I) and (II).

The compounds of Formulas (I) and (II) are useful as immune responsemodifiers due to their ability to induce cytokine biosynthesis andotherwise modulate the immune response when administered to animals.This makes the compounds useful in the treatment of a variety ofconditions such as viral diseases and tumors that are responsive to suchchanges in the immune response.

The invention further provides pharmaceutical compositions containingthe immune response modifying compounds, and methods of inducingcytokine biosynthesis in an animal, treating a viral infection in ananimal, and/or treating a neoplastic disease in an animal byadministering a compound of Formula (I) or (II) to the animal.

In addition, the invention provides methods of synthesizing thecompounds of the invention and novel intermediates useful in thesynthesis of these compounds.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned earlier, we have found certain compounds that inducecytokine biosynthesis and modify the immune response in animals. Suchcompounds are represented by Formulas (I) and (II) as shown below.

Imidazoquinoline compounds of the invention, which have ether and amidefunctionality at the 1-position, are represented by Formula (I):

wherein:

X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-;

R₁ is selected from the group consisting of:

—R₄—CR₃—Z—R₆-alkyl;

—R₄—CR₃—Z—R₆-alkenyl;

—R₄—CR₃—Z—R₆-aryl;

—R₄—CR₃—Z—R₆-heteroaryl;

—R₄—CR₃—Z—R₆-heterocyclyl;

—R₄—CR₃—Z—H;

—R₄—NR₇—CR₃—R₆-alkyl;

—R₄—NR₇—CR₃—R₆-alkenyl;

R₄—NR₇—CR₃—R₆-aryl;

—R₄—NR₇—CR₃—R₆-heteroaryl;

—R₄—NR₇CR₃—R₆-heterocyclyl; and

—R₄—NR₇—CR₃—R₈;

each Z is independently —NR₅—, —O—, or —S—;

R₂ is selected from the group consisting of:

-hydrogen;

-alkyl;

-alkenyl;

-aryl;

-heteroaryl;

-heterocyclyl;

-alkyl-Y-alkyl;

-alkyl-Y-alkenyl;

-alkyl-Y-aryl; and

-alkyl or alkenyl substituted by one or more substituents selected fromthe group consisting of:

—OH;

-halogen;

—N(R₅)₂;

—CO—N(R₅)₂;

—CO—C₁₋₁₀ alkyl;

—CO—O—C₁₋₁₀ alkyl;

—N₃;

-aryl;

-heteroaryl;

-heterocyclyl;

—CO-aryl; and

—CO-heteroaryl;

each R₃ is ═O or ═S;

each R₄ is independently alkyl or alkenyl, which may be interrupted byone or more —O— groups;

each R₅ is independently H or C₁₋₁₀ alkyl;

R₆ is a bond, alkyl, or alkenyl, which may be interrupted by one or more—O— groups;

R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or R₄ and R₇ can join together toform a ring;

R₈ is H or C₁₋₁₀ alkyl; or R₇ and R₈ can join together to form a ring;

each Y is independently —O— or —S(O)₀₋₂—;

n is 0 to 4; and

each R present is independently selected from the group consisting ofC₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen and trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

The invention also includes tetrahydroimidazoquinoline compounds thatbear an ether and amide containing substituent at the 1-position. Suchtetrahydroimidazoquinoline compounds are represented by Formula (II):

wherein:

X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-;

R₁ is selected from the group consisting of:

—R₄—CR₃—Z—R₆-alkyl;

—R₄—R₃—Z—R₆-alkenyl;

—R₄—CR₃—Z—R₆-aryl;

—R₄—CR₃—Z—R₆-heteroaryl;

—R₄—CR₃—Z—R₆-heterocyclyl;

—R₄—CR₃—Z—H;

—R₄—NR₇—CR₃—R₆-alkyl;

—R₄—NR₇—CR₃—R₆-alkenyl;

—R₄—NR₇—CR₃—R₆-aryl;

—R₄—NR₇—CR₃—R₆-heteroaryl;

—R₄—NR₇—CR₃—R₆-heterocyclyl;

—R₄—NR₇—CR₃—R₈;

each Z is independently —NR₅—, —O—, or —S—;

R₂ is selected from the group consisting of:

-hydrogen;

-alkyl;

-alkenyl;

-aryl;

-heteroaryl;

-heterocyclyl;

-alkyl-Y-alkyl;

-alkyl-Y-alkenyl;

-alkyl-Y-aryl; and

-alkyl or alkenyl substituted by one or more substituents selected fromthe group consisting of:

—OH;

-halogen;

—N(R₅)₂;

—CO—N(R₅)₂;

—CO—C₁₋₁₀ alkyl;

—CO—C₁₋₁₀ alkyl;

—N₃;

-aryl;

-heteroaryl;

-heterocyclyl;

—CO-aryl; and

—CO-heteroaryl;

each R₃ is ═O or ═S;

each R₄ is independently alkyl or alkenyl, which may be interrupted byone or more —O— groups;

each R₅ is independently H or C₁₋₁₀ alkyl;

R₆ is a bond, alkyl, or alkenyl, which may be interrupted by one or more—O— groups;

R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or R₄ and R₇ can join together toform a ring;

R₈ is H or C₁₋₁₀ alkyl; or R₇ and R₈ can join together to form a ring;

each Y is independently —O— or —S(O)₀₋₂—;

n is 0 to 4; and

each R present is independently selected from the group consisting ofC₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen, and trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

Preparation of the Compounds

Compounds of the invention can be prepared according to Reaction SchemeI where R, R₂, R₃, R₄, X, Z and n are as defined above and R₁₁ is—R₆-alkyl, —R₆-aryl, —R₆-heteroaryl or —R₆-heterocyclyl where R₆ is asdefined above.

In step (1) of Reaction Scheme I a 1H-imidazo[4,5-c]quinolin-1-ylalcohol of Formula X is alkylated with a halide of Formula XI to providea 1H-imidazo[4,5-c]quinolin-1-yl ether of Formula XII. The alcohol ofFormula X is reacted with sodium hydride in a suitable solvent such asN,N-dimethylformamide to form an alkoxide. Alternatively, the alkoxidecan be formed by adding the alcohol to a biphasic mixture of aqueous 50%sodium hydroxide and an inert solvent such as dichloromethane in thepresence of a phase transfer catalyst such as benzyltrimethylammoniumchloride. The alkoxide is then combined with the halide. The reactioncan be carried out at ambient temperature. Many compounds of Formula Xare known, see for example, Gerster, U.S. Pat. No. 4,689,338; others canreadily be prepared using known synthetic routes, see for example,Gerster et al., U.S. Pat. No. 5,605,899 and Gerster, U.S. Pat. No.5,175,296. Many halides of Formula XI are commercially available; otherscan be readily prepared using known synthetic routes.

In step (2) of Reaction Scheme I a 1H-imidazo[4,5-c]quinolin-1-yl etherof Formula XII is oxidized to provide a1H-imidazo[4,5-c]quinoline-5N-oxide of Formula XIII using a conventionaloxidizing agent capable of forming N-oxides. Preferably a solution of acompound of Formula XII in chloroform is oxidized using3-chloroperoxybenzoic acid at ambient temperature.

In step (3) of Reaction Scheme I a 1H-imidazo[4,5-c]quinoline-5N-oxideof Formula XIII is aminated to provide a1H-imidazo[4,5-c]quinolin-4-amine of Formula XIV which is a subgenus ofFormula I. Step (3) involves (i) reacting a compound of Formula XIIIwith an acylating agent and then (ii) reacting the product with anaminating agent. Part (i) of step (3) involves reacting an N-oxide ofFormula XIII with an acylating agent. Suitable acylating agents includealkyl- or arylsulfonyl chlorides (e.g., benezenesulfonyl chloride,methanesulfonyl chloride, p-toluenesulfonyl chloride). Arylsulfonylchlorides are preferred. Para-toluenesulfonyl chloride is mostpreferred. Part (ii) of step (3) involves reacting the product of part(i) with an excess of an aminating agent. Suitable aminating agentsinclude ammonia (e.g., in the form of ammonium hydroxide) and ammoniumsalts (e.g., ammonium carbonate, ammonium bicarbonate, ammoniumphosphate). Ammonium hydroxide is preferred. The reaction is preferablycarried out by dissolving the N-oxide of Formula XIII in an inertsolvent such as dichloromethane, adding the aminating agent to thesolution, and then slowly adding the acylating agent. The product or apharmaceutically acceptable salt thereof can be isolated usingconventional methods.

Alternatively, step (3) may be carried out by (i) reacting an N-oxide ofFormula XIII with an isocyanate and then (ii) hydrolyzing the resultingproduct. Part (i) involves reacting the N-oxide with an isocyanatewherein the isocyanato group is bonded to a carbonyl group. Preferredisocyanates include trichloroacetyl isocyanate and aroyl isocyanatessuch as benzoyl isocyanate. The reaction of the isocyanate with theN-oxide is carried out under substantially anhydrous conditions byadding the isocyanate to a solution of the N-oxide in an inert solventsuch as chloroform or dichloromethane. Part (ii) involves hydrolysis ofthe product from part (i). The hydrolysis can be carried out byconventional methods such as heating in the presence of water or a loweralkanol optionally in the presence of a catalyst such as an alkali metalhydroxide or lower alkoxide. The product or a pharmaceuticallyacceptable salt thereof can be isolated using conventional methods.

Compounds of the invention can be prepared according to Reaction SchemeII where R, R₂, R₄, R₇, R₁₁, X and n are as defined above and BOC istert-butoxycarbonyl.

In step (1) of Reaction Scheme II the amino group of an aminoalcohol ofFormula XV is protected with a tert-butoxycarbonyl group. A solution ofthe aminoalcohol in tetrahydrofuran is treated with di-tert-butyldicarbonate in the presence of a base such as sodium hydroxide. Manyaminoalcohols of Formula XV are commercially available; others can beprepared using known synthetic methods.

In step (2) of Reaction Scheme II a protected aminoalcohol of FormulaXVI is converted to an iodide of Formula XVII. Iodine is added to asolution of triphenylphosphine and imidazole in dichloromethane; then asolution of a protected aminoalcohol of Formula XVI in dichloromethaneis added. The reaction is carried out at ambient temperature.

In step (3) of Reaction Scheme II a 1H-imidazo[4,5-c]quinolin-1-ylalcohol of Formula X is alkylated with an iodide of Formula XVII toprovide a 1H-imidazo[4,5-c]quinolin-1-yl ether of Formula XVIII. Thealcohol of Formula X is reacted with sodium hydride in a suitablesolvent such as N,N-dimethylformamide to form an alkoxide. The iodide isadded to the alkoxide solution at ambient temperature. After theaddition is complete the reaction is stirred at an elevated temperature(˜100° C.).

In step (4) of Reaction Scheme II a 1H-imidazo[4,5-c]quinolin-1-yl etherof Formula XVIII is oxidized to provide a1H-imidazo[4,5-c]quinoline-5N-oxide of Formula XIX using a conventionaloxidizing agent capable of forming N-oxides. Preferably a solution of acompound of Formula XVIII in chloroform is oxidized using3-chloroperoxybenzoic acid at ambient temperature.

In step (5) of Reaction Scheme II a 1H-imidazo[4,5-c]quinoline-5N-oxideof Formula XIX is aminated to provide a1H-imidazo[4,5-c]quinolin-4-amine of Formula XX. Step (5) involves (i)reacting a compound of Formula XIX with an acylating agent and then (ii)reacting the product with an aminating agent. Part (i) of step (5)involves reacting an N-oxide of Formula XIX with an acylating agent.Suitable acylating agents include alkyl- or arylsulfonyl chlorides(e.g., benezenesulfonyl chloride, methanesulfonyl chloride,p-toluenesulfonyl chloride). Arylsulfonyl chlorides are preferred.Para-toluenesulfonyl chloride is most preferred. Part (ii) of step (5)involves reacting the product of part (i) with an excess of an aminatingagent. Suitable aminating agents include ammonia (e.g., in the form ofammonium hydroxide) and ammonium salts (e.g., ammonium carbonate,ammonium bicarbonate, ammonium phosphate). Ammonium hydroxide ispreferred. The reaction is preferably carried out by dissolving theN-oxide of Formula XIX in an inert solvent such as dichloromethane or1,2-dichloroethane with heating if necessary, adding the aminating agentto the solution, and then slowly adding the acylating agent. Optionallythe reaction can be carried out in a sealed pressure vessel at anelevated temperature (85-100°).

In step (6) of Reaction Scheme II the protecting group is removed byhydrolysis under acidic conditions to provide a1H-imidazo[4,5-c]quinolin-4-amine of Formula XXI. Preferably thecompound of Formula XX is treated with hydrochloric acid/ethanol atambient temperature or with gentle heating.

In step (7) of Reaction Scheme II a 1H-imidazo[4,5-c]quinolin-4-amine ofFormula XXI is converted to an amide of Formula XXII which is a subgenusof Formula I using conventional synthetic methods. For example, acompound of Formula XXI can be reacted with an acid chloride of FormulaR₁₁C(O)Cl. The reaction can be carried out by adding a solution of theacid chloride in a suitable solvent such as dichloromethane or1-methyl-2-pyrrolidinone to a solution of a compound of Formula XXI atambient temperature. Alternatively, a compound of Formula XXI can bereacted with an acid of Formula R₁₁COOH. The reaction can be carried outat ambient temperature in a solvent such as dichloromethane or pyridineusing a standard coupling reagent such as 1,3-dicyclohexylcarbodiimideor 1[3-(dimethylamino)propyl]-3-ethylcarbodiimide. The product or apharmaceutically acceptable salt thereof can be isolated usingconventional methods.

Compounds of the invention can be prepared according to Reaction SchemeIII where R, R₂, R₄, R₇, R₁₁, X and n are as defined above and BOC istert-butoxycarbonyl.

In step (1) of Reaction Scheme III the amino group of an aminoalcohol ofFormula XXIII is protected with a tert-butoxycarbonyl group. A solutionof the aminoalcohol in tetrahydrofuran is treated with di-tert-butyldicarbonate in the presence of a base such as sodium hydroxide. Manyaminoalcohols of Formula XXIII are commercially available; others can beprepared using known synthetic methods.

In step (2) of Reaction Scheme III a protected amino alcohol of FormulaXXIV is converted to a methanesulfonate of Formula XXV. A solution of acompound of Formula XXIV in a suitable solvent such as dichloromethaneis treated with methanesulfonyl chloride in the presence of a base suchas triethylamine. The reaction can be carried out at a reducedtemperature (0°).

In step (3a) of Reaction Scheme III a methanesulfonate of Formula XXV isconverted to an azide of Formula XXVI. Sodium azide is added to asolution of a compound of Formula XXV in a suitable solvent such asN,N-dimethylformamide. The reaction can be carried out at an elevatedtemperature (80-100° C.).

In step (3b) of Reaction Scheme III a compound of Formula XXVI isalkylated with a halide of Formula Hal-R₇ to provide a compound ofFormula XXVII. In compounds where R₇ is hydrogen this step is omitted.The compound of Formula XXVI is reacted with sodium hydride in asuitable solvent such as N,N-dimethylformamide to form the anion andthen combined with the halide. The reaction can be carried out atambient temperature.

In step (4) of Reaction Scheme III an azide of Formula XXVI or XXVII isreduced to provide an amine of Formula XXVIII. Preferably, the reductionis carried out using a conventional heterogeneous hydrogenation catalystsuch as palladium on carbon. The reaction can conveniently be carriedout on a Parr apparatus in a suitable solvent such as methanol orisopropanol.

In step (5) of Reaction Scheme III a 4-chloro-3-nitroquinoline ofFormula XXIX is reacted with an amine of Formula XXVIII to provide a3-nitroquinoline of Formula XXX. The reaction can be carried out byadding an amine of Formula XXVIII to a solution of a compound of FormulaXXIX in a suitable solvent such as dichloromethane in the presence of abase such as triethylamine. Many quinolines of Formula XXIX are knowncompounds or can be prepared using known synthetic methods, see forexample, U.S. Pat. No. 4,689,338 and references cited therein.

In step (6) of Reaction Scheme III a 3-nitroquinoline of Formula XXX isreduced to provide a 3-aminoquinoline of Formula XXXI. Preferably, thereduction is carried out using a conventional heterogeneoushydrogenation catalyst such as platinum on carbon. The reaction canconveniently be carried out on a Parr apparatus in a suitable solventsuch as toluene.

In step (7) of Reaction Scheme III a compound of Formula XXXI is reactedwith a carboxylic acid or an equivalent thereof to provide a1H-imidazo[4,5-c]quinoline of Formula XVIII. Suitable equivalents tocarboxylic acid include orthoesters, and 1,1-dialkoxyalkyl alkanoates.The carboxylic acid or equivalent is selected such that it will providethe desired R₂ substituent in a compound of Formula XVIII. For example,triethyl orthoformate will provide a compound where R₂ is hydrogen andtriethyl orthovalerate will provide a compound where R₂ is butyl. Thereaction can be run in the absence of solvent or in an inert solventsuch as toluene. The reaction is run with sufficient heating to driveoff any alcohol or water formed as a byproduct of the reaction.Optionally a catalyst such as pyridine hydrochloride can be included.

Alternatively, step (7) can be carried out by (i) reacting a compound ofFormula XXXI with an acyl halide of Formula R₂C(O)Cl or R₂C(O)Br andthen (ii) cyclizing. In part (i) the acyl halide is added to a solutionof a compound of Formula XXXI in an inert solvent such as acetonitrileor dichloromethane. The reaction can be carried out at ambienttemperature or at a reduced temperature. In part (ii) the product ofpart (i) is heated in an alcoholic solvent in the presence of a base.Preferably the product of part (i) is refluxed in ethanol in thepresence of an excess of triethylamine or heated with methanolicammonia.

Steps (8), (9), (10) and (11) are carried out in the same manner assteps (4), (5), (6) and (7) of Reaction Scheme II.

Compounds of the invention can be prepared according to Reaction SchemeIV where R, R₁, R₂, X and n are as defined above In Reaction Scheme IV a4-amino-1H-imidazo[4,5-c]quinolin-1-yl alcohol of Formula XXXII isalkylated with a halide of Formula XXXIII to provide a1H-imidazo[4,5-c]quinolin-4-amine of Formula I. The alcohol of FormulaXXXII is reacted with sodium hydride in a suitable solvent such asN,N-dimethylformamide to form an alkoxide. The halide is then added tothe reaction mixture. The reaction can be carried out at ambienttemperature or with gentle heating (˜50° C.) if desired. The product ora pharmaceutically acceptable salt thereof can be isolated usingconventional methods.

Many compounds of Formula XXXII are known, see for example Gerster, U.S.Pat. No. 4,689,338 and Gerster et. al., U.S. Pat. No. 5,605,899, thedisclosures of which are incorporated by reference herein; others canreadily be prepared using known synthetic routes, see for example, Andreet. al, U.S. Pat. No. 5,578,727; Gerster, U.S. Pat. No. 5,175,296;Nikolaides et al., U.S. Pat. No. 5,395,937; and Gerster et. al., U.S.Pat. No. 5,741,908, the disclosures of which are incorporated byreference herein. Many halides of Formula XXXIII are commerciallyavailable; others can be readily prepared using known synthetic methods.

Compounds of the invention can be prepared according to Reaction SchemeV where R, R₂, R₄, R₇, R₁₁, X and n are as defined above.

In step (1) of Reaction Scheme V a 1H-imidazo[4,5-c]quinolin-4-amine ofFormula XXI is reduced to provide a6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine of Formula XXXIV.Preferably the reduction is carried out by suspending or dissolving acompound of Formula XXI in trifluoroacetic acid, adding a catalyticamount of platinum (IV) oxide, and then hydrogenating. The reaction canbe conveniently carried out in a Parr apparatus.

Step (2) is carried out in the same manner as step (7) of ReactionScheme II to provide a6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine of Formula XXXVwhich is a subgenus of Formula II. The product or a pharmaceuticallyacceptable salt thereof can be isolated using conventional methods.

Compounds of the invention can be prepared according to Reaction SchemeVI where R, R₁, R₂, X and n are as defined above.

In Reaction Scheme VI a4-amino-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-1-yl alcohol ofFormula XXXVI is alkylated with a halide of Formula XXXIII to provide a6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine of Formula II. Thealcohol of Formula XXXVI is reacted with sodium hydride in a suitablesolvent such as N,N-dimethylformamide to form an alkoxide. The halide isthen added to the reaction mixture. The reaction can be carried out atambient temperature or with gentle heating (˜50° C.) if desired. Theproduct or a pharmaceutically acceptable salt thereof can be isolatedusing conventional methods.

Many 6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolines of Formula XXXVI areknown, see for example, Nikolaides et al., U.S. Pat. No. 5,352,784;others can be prepared using known synthetic methods, see for example,Lindstrom, U.S. Pat. No. 5,693,811; the disclosures of which areincorporated by reference herein.

The invention also provides novel compounds useful as intermediates inthe synthesis of the compounds of Formulas (I) and (II). Theseintermediate compounds have the structural Formulas (III)-(V), describedin more detail below.

One class of intermediate compounds has Formula (III):

wherein:

X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-;

R₁ is selected from the group consisting of:

—R₄—CR₃—Z—R₆-alkyl;

—R₄—CR₃—Z—R₆-alkenyl;

—R₄—CR₃—Z—R₆-aryl;

—R₄—CR₃—Z—R₆-heteroaryl;

—R₄—CR₃—Z—R₆-heterocyclyl;

—R₄—CR₃—Z—H;

—R₄—NR₇—CR₃—R₆-alkyl;

—R₄—NR₇—CR₃—R₆-alkenyl;

—R₄—NR₇CR₃—R₆-aryl;

—R₄—NR₇—CR₃—R₆-heteroaryl;

—R₄—NR₇—CR₃—R₆-heterocyclyl; and

—R₄—NR₇—CR₃—R₈;

each Z is independently —NR₅—, —O—, or —S—;

R₂ is selected from the group consisting of:

-hydrogen;

-alkyl;

-alkenyl;

-aryl;

-heteroaryl;

-heterocyclyl;

-alkyl-Y-alkyl;

-alkyl-Y-alkenyl;

-alkyl-Y-aryl; and

-alkyl or alkenyl substituted by one or more substituents selected fromthe group consisting of:

—OH;

-halogen;

—N(R₅)₂;

—CO—N(R₅)₂;

—CO—C₁₋₁₀ alkyl;

—CO—O—C₁₋₁₀ alkyl;

—N₃;

-aryl;

-heteroaryl;

-heterocyclyl;

—CO-aryl; and

—CO-heteroaryl;

each R₃ is ═O or ═S;

each R₄ is independently alkyl or alkenyl, which may be interrupted byone or more —O— groups;

each R₅ is independently H or C₁₋₁₀ alkyl;

R₆ is a bond, or is alkyl, or alkenyl, which may be interrupted by oneor more —O— groups;

R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or R₄ and R₇ can join to form aring;

R₈ is H or C₁₋₁₀ alkyl; or R₇ and R₈ can join to form a ring;

each Y is independently —O— or —S(O)₀₋₂—;

n is 0 to 4; and

each R present is independently selected from the group consisting ofC₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen and trifluoromethyl; or apharmaceutically acceptable salt thereof.

Another class of intermediates is described by formula (IV):

wherein:

X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-;

R₁ is selected from the group consisting of:

—R₄—CR₃—Q—R₆-alkyl;

—R₄—CR₃—Q—R₆-alkenyl;

R—CR₃—Q—R₆-aryl;

—R₄—CR₃—Q—R₆-heteroaryl;

—R₄—CR₃—Q—R₆-heterocyclyl;

—R₄—CR₃—Q—H;

—R₄—NR₅—CR₃—R₆-alkyl;

—R₄—NR₅—CR₃—R₆-alkenyl;

—R₄—NR₇—CR₃—R₆-aryl;

—R₄—NR₇—CR₃—R₆-heteroaryl;

—R₄—NR₇—CR₃—R₆-heterocyclyl; and

—R—NR₅—CR₃—R₈;

each Q is independently —NR₅— or —O—;

each R₃ is ═O or ═S;

each R₄ is independently alkyl or alkenyl, which may be interrupted byone or more —O— groups;

each R₅ is independently H or C₁₋₁₀ alkyl;

R₆ is a bond, alkyl, or alkenyl, which may be interrupted by one or more—O— groups;

R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or R₄ and R₇ can join to form aring;

R₈ is H or C₁₋₁₀ alkyl; or R₄ and R₇ can join to form a ring;

n is 0 to 4; and

each R present is independently selected from the group consisting ofC₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, halogen and trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

An additional class of intermediate compounds has the formula (V):

wherein:

X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-;

R₂ is selected from the group consisting of:

-hydrogen;

-alkyl;

-alkenyl;

-aryl;

-heteroaryl;

-heterocyclyl;

-alkyl-Y-alkyl;

-alkyl-Y-alkenyl;

-alkyl-Y-aryl; and

-alkyl or alkenyl substituted by one or more substituents selected fromthe group consisting of:

—OH;

-halogen;

—N(R₅)₂;

—CO—N(R₅)₂;

—CO—C₁₋₁₀ alkyl;

—CO—O—C₁₋₁₀ alkyl;

—N₃;

-aryl;

-heteroaryl;

-heterocyclyl;

—CO-aryl; and

—CO-heteroaryl;

each R₄ is independently alkyl or alkenyl, which may be interrupted byone or more —O— groups;

R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or R₄ and R₇ can join to form aring;

each Y is independently —O— or —S(O)₀₋₂—;

n is 0 to 4; and

each R present is independently selected from the group consisting ofC₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen and trifluoromethyl;

or a pharmaceutically acceptable salt thereof.

As used herein, the terms “alkyl”, “alkenyl” and the prefix “alk-” areinclusive of both straight chain and branched chain groups and of cyclicgroups, i.e. cycloalkyl and cycloalkenyl. Unless otherwise specified,these groups contain from 1 to 20 carbon atoms, with alkenyl groupscontaining from 2 to 20 carbon atoms. Preferred groups have a total ofup to 10 carbon atoms. Cyclic groups can be monocyclic or polycyclic andpreferably have from 3 to 10 ring carbon atoms. Exemplary cyclic groupsinclude cyclopropyl, cyclopropylmethyl, cyclopentyl, cyclohexyl andadamantyl.

In addition, the alkyl and alkenyl portions of-X-groups can beunsubstituted or substituted by one or more substituents, whichsubstituents are selected from the groups consisting of alkyl, alkenyl,aryl, heteroaryl, heterocyclyl, arylalkyl, heteroarylalkyl, andheterocyclylalkyl.

The term “haloalkyl” is inclusive of groups that are substituted by oneor more halogen atoms, including perfluorinated groups. This is alsotrue of groups that include the prefix “halo-”. Examples of suitablehaloalkyl groups are chloromethyl, trifluoromethyl, and the like.

The term “aryl” as used herein includes carbocyclic aromatic rings orring systems. Examples of aryl groups include phenyl, naphthyl,biphenyl, fluorenyl and indenyl. The term “heteroaryl” includes aromaticrings or ring systems that contain at least one ring hetero atom (e.g.,O, S, N). Suitable heteroaryl groups include furyl, thienyl, pyridyl,quinolinyl, isoquinolinyl, indolyl, isoindolyl, triazolyl, pyrrolyl,tetrazolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, benzofuranyl,benzothiophenyl, carbazolyl, benzoxazolyl, pyrimidinyl, benzimidazolyl,quinoxalinyl, benzothiazolyl, naphthyridinyl, isoxazolyl, isothiazolyl,purinyl, quinazolinyl, and so on.

“Heterocyclyl” includes non-aromatic rings or ring systems that containat least one ring hetero atom (e.g., O, S, N) and includes all of thefully saturated and partially unsaturated derivatives of the abovementioned heteroaryl groups. Exemplary heterocyclic groups includepyrrolidinyl, tetrahydrofuranyl, morpholinyl, thiomorpholinyl,piperidinyl, piperazinyl, thiazolidinyl, imidazolidinyl,isothiazolidinyl, and the like.

The aryl, heteroaryl, and heterocyclyl groups can be unsubstituted orsubstituted by one or more substituents independently selected from thegroup consisting of alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy,haloalkylthio, halogen, nitro, hydroxy, mercapto, cyano, carboxy,formyl, aryl, aryloxy, arylthio, arylalkoxy, arylalkylthio, heteroaryl,heteroaryloxy, heteroarylthio, heteroarylalkoxy, heteroarylalkylthio,amino, alkylamino, dialkylamino, heterocyclyl, heterocycloalkyl,alkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, haloalkylcarbonyl,haloalkoxycarbonyl, alkylthiocarbonyl, arylcarbonyl, heteroarylcarbonyl,aryloxycarbonyl, heteroaryloxycarbonyl, arylthiocarbonyl,heteroarylthiocarbonyl, alkanoyloxy, alkanoylthio, alkanoylamino,aroyloxy, aroylthio, aroylamino, alkylaminosulfonyl, alkylsulfonyl,arylsulfonyl, heteroarylsulfonyl, aryldiazinyl, alkylsulfonylamino,arylsulfonylamino, arylalkylsulfonylamino, alkylcarbonylamino,alkenylcarbonylamino, arylcarbonylamino, arylalkylcarbonylamino,heteroarylcarbonylamino, heteroarylalkycarbonylamino,alkylsulfonylamino, alkenylsulfonylamino, arylsulfonylamino,arylalkylsulfonylamino, heteroarylsulfonylamino,heteroarylalkylsulfonylamino, alkylaminocarbonylamino,alkenylaminocarbonylamino, arylaminocarbonylamino,arylalkylaminocarbonylamino, heteroarylaminocarbonylamino,heteroarylalkylaminocarbonylamino and, in the case of heterocyclyl, oxo.If any other groups are identified as being “substituted” or “optionallysubstituted”, then those groups can also be substituted by one or moreof the above enumerated substituents.

Certain substituents are generally preferred. For example, preferred R₁groups include —R₄—CR₃—Z—R₆-alkyl and —R₄—CR₃—Z—R₆-aryl, wherein thealkyl and aryl groups can be unsubstituted or substituted; R₃ ispreferably ═O; R₄ is preferably ethylene or n-butylene; and Z ispreferably —NR₅—. Preferably no R substituents are present (i.e., n is0). Preferred R₂ groups include alkyl groups having 1 to 4 carbon atoms(i.e., methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl,and cyclopropylmethyl), methoxyethyl, and ethoxymethyl. For substitutedgroups such as substituted alkyl or substituted aryl groups, preferredsubstituents include halogen, nitrile, methoxy, trifluoromethyl, andtrifluoromethoxy. One or more of these preferred substituents, ifpresent, can be present in the compounds of the invention in anycombination.

The invention is inclusive of the compounds described herein in any oftheir pharmaceutically acceptable forms, including isomers (e.g.,diastereomers and enantiomers), salts, solvates, polymorphs, and thelike. In particular, if a compound is optically active, the inventionspecifically includes each of the compound's enantiomers as well asracemic mixtures of the enantiomers.

Pharmaceutical Compositions and Biological Activity

Pharmaceutical compositions of the invention contain a therapeuticallyeffective amount of a compound of the invention as described above incombination with a pharmaceutically acceptable carrier.

The term “a therapeutically effective amount” means an amount of thecompound sufficient to induce a therapeutic effect, such as cytokineinduction, antitumor activity, and/or antiviral activity. Although theexact amount of active compound used in a pharmaceutical composition ofthe invention will vary according to factors known to those of skill inthe art, such as the physical and chemical nature of the compound, thenature of the carrier, and the intended dosing regimen, it isanticipated that the compositions of the invention will containsufficient active ingredient to provide a dose of about 100 ng/kg toabout 50 mg/kg, preferably about 10 μg/kg to about 5 mg/kg, of thecompound to the subject. Any of the conventional dosage forms may beused, such as tablets, lozenges, parenteral formulations, syrups,creams, ointments, aerosol formulations, transdermal patches,transmucosal patches and the like.

The compounds of the invention can be administered as the singletherapeutic agent in the treatment regimen, or the compounds of theinvention may be administered in combination with one another or withother active agents, including additional immune response modifiers,antivirals, antibiotics, etc.

The compounds of the invention have been shown to induce the productionof certain cytokines in experiments performed according to the tests setforth below. These results indicate that the compounds are useful asimmune response modifiers that can modulate the immune response in anumber of different ways, rendering them useful in the treatment of avariety of disorders.

Cytokines whose production may be induced by the administration ofcompounds according to the invention generally include interferon-α(IFN-α) and/or tumor necrosis factor-cc (TNF-α) as well as certaininterleukins (IL). Cytokines whose biosynthesis may be induced bycompounds of the invention include IFN-α, TNF-α, IL-1, IL-6, IL-10 andIL-12, and a variety of other cytokines. Among other effects, these andother cytokines can inhibit virus production and tumor cell growth,making the compounds useful in the treatment of viral diseases andtumors. Accordingly, the invention provides a method of inducingcytokine biosynthesis in an animal comprising administering an effectiveamount of a compound or composition of the invention to the animal.

Certain compounds of the invention have been found to preferentiallyinduce the expression of IFN-α in a population of hematopoietic cellssuch as PBMCs (peripheral blood mononuclear cells) containing pDC2 cells(precursor dendritic cell-type 2) without concomitant production ofsignificant levels of inflammatory cytokines.

In addition to the ability to induce the production of cytokines, thecompounds of the invention affect other aspects of the innate immuneresponse. For example, natural killer cell activity may be stimulated,an effect that may be due to cytokine induction. The compounds may alsoactivate macrophages, which in turn stimulates secretion of nitric oxideand the production of additional cytokines. Further, the compounds maycause proliferation and differentiation of B-lymphocytes.

Compounds of the invention also have an effect on the acquired immuneresponse. For example, although there is not believed to be any directeffect on T cells or direct induction of T cell cytokines, theproduction of the T helper type 1 (Th1) cytokine IFN-γ is inducedindirectly and the production of the T helper type 2 (Th2) cytokinesIL-4, IL-5 and IL-13 are inhibited upon administration of the compounds.This activity means that the compounds are useful in the treatment ofdiseases where upregulation of the Th1 response and/or downregulation ofthe Th2 response is desired. In view of the ability of compounds of theinvention to inhibit the Th2 immune response, the compounds are expectedto be useful in the treatment of atopic diseases, e.g., atopicdermatitis, asthma, allergy, allergic rhinitis; systemic lupuserythematosis; as a vaccine adjuvant for cell mediated immunity; andpossibly as a treatment for recurrent fungal diseases and chlamydia.

The immune response modifying effects of the compounds make them usefulin the treatment of a wide variety of conditions. Because of theirability to induce the production of cytokines such as IFN-α and/orTNF-α, the compounds are particularly useful in the treatment of viraldiseases and tumors. This immunomodulating activity suggests thatcompounds of the invention are useful in treating diseases such as, butnot limited to, viral diseases including genital warts; common warts;plantar warts; Hepatitis B; Hepatitis C; Herpes Simplex Virus Type I andType II; molluscum contagiosum; varriola major; HIV; CMV; VZV;rhinovirus; adenovirus; influenza; and para-influenza; intraepithelialneoplasias such as cervical intraepithelial neoplasia; humanpapillomavirus (HPV) and associated neoplasias; fungal diseases, e.g.candida, aspergillus, and cryptococcal meningitis; neoplastic diseases,e.g., basal cell carcinoma, hairy cell leukemia, Kaposi's sarcoma, renalcell carcinoma, squamous cell carcinoma, myelogenous leukemia, multiplemyeloma, melanoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma,and other cancers; parasitic diseases, e.g. pneumocystis carnii,cryptosporidiosis, histoplasmosis, toxoplasmosis, trypanosome infection,and leishmaniasis; and bacterial infections, e.g., tuberculosis, andmycobacterium avium. Additional diseases or conditions that can betreated using the compounds of the invention include actinic keratosis;eczema; eosinophilia; essential thrombocythaemia; leprosy; multiplesclerosis; Ommen's syndrome; discoid lupus; Bowen's disease; Bowenoidpapulosis; alopecia areata; the inhibition of Keloid formation aftersurgery and other types of post-surgical scars. In addition, thesecompounds could enhance or stimulate the healing of wounds, includingchronic wounds. The compounds may be useful for treating theopportunistic infections and tumors that occur after suppression of cellmediated immunity in, for example, transplant patients, cancer patientsand HIV patients.

An amount of a compound effective to induce cytokine biosynthesis is anamount sufficient to cause one or more cell types, such as monocytes,macrophages, dendritic cells and B-cells to produce an amount of one ormore cytokines such as, for example, IFN-α, TNF-α, IL-1, IL-6, IL-10 andIL-12 that is increased over the background level of such cytokines. Theprecise amount will vary according to factors known in the art but isexpected to be a dose of about 100 ng/kg to about 50 mg/kg, preferablyabout 10 μg/kg to about 5 mg/kg. The invention also provides a method oftreating a viral infection in an animal and a method of treating aneoplastic disease in an animal comprising administering an effectiveamount of a compound or composition of the invention to the animal. Anamount effective to treat or inhibit a viral infection is an amount thatwill cause a reduction in one or more of the manifestations of viralinfection, such as viral lesions, viral load, rate of virus production,and mortality as compared to untreated control animals. The preciseamount will vary according to factors known in the art but is expectedto be a dose of about 100 ng/kg to about 50 mg/kg, preferably about 10μg/kg to about 5 mg/kg. An amount of a compound effective to treat aneoplastic condition is an amount that will cause a reduction in tumorsize or in the number of tumor foci. Again, the precise amount will varyaccording to factors known in the art but is expected to be a dose ofabout 100 ng/kg to about 50 mg/kg, preferably about 10 μg/kg to about 5mg/kg.

The invention is further described by the following examples, which areprovided for illustration only and are not intended to be limiting inany way.

In the examples below some of the compounds were purified usingsemi-preparative HPLC. Two different methods were used and they aredescribed below.

Method A

This method used a A-100 Gilson-6 equipped with 900 Series IntelligentInterface. The semi-prep HPLC fractions were analyzed by LC-APCI/MS andthe appropriate fractions were combined and lyophilized to provide thetrifluoroacetate salt of the desired compound.

Column: column Microsorb C18, 21.4×250 mm, 8 micron particle size, 60 Åpore; flow rate: 10 mL/min.; gradient elution from 2-95% B in 25 min.,hold at 95% B for 5 min., where A=0.1% trifluoroacetic acid/water andB=0. 1% trifluoroacetic acid/acetonitrile; peak detection at 254 nm fortriggering fraction collection.

Method B

This method used a Waters Fraction Lynx automated purification system.The semi-prep HPLC fractions were analyzed using a Micromass LC-TOFMSand the appropriate fractions were combined and centrifuge evaporated toprovide the trifluoroacetate salt of the desired compound. The structurewas confirmed by ¹H NMR spectroscopy.

Column: Phenomenex Luna C18(2), 10×50 mm, 5 micron particle size, 100 Åpore; flow rate: 25 mL/min.; gradient elution from 5-65% B in 4 min.,then 65 to 95% B in 0.1 min, then hold at 95% B for 0.4 min., whereA=0.05% trifluoroacetic acid/water and B=0.05% trifluoroaceticacid/acetonitrile; fraction collection by mass-selective triggering.

EXAMPLE 11-[2-(2-aminoethoxy)ethyl]-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine

Part A

A solution of 2-(2-aminoethoxy)ethanol (29.0 g, 0.276 mol) in 180 mL oftetrahydrofuran (THF), under N₂, was cooled to 0° C. and treated with140 mL of 2N NaOH solution. A solution of di-tert-butyl dicarbonate(60.2 g, 0.276 mol) in 180 mL of THF was then added dropwise over 1 h tothe rapidly stirred solution. The reaction mixture was then allowed towarm to room temperature and was stirred an additional 18 h. The THF wasthen removed under reduced pressure and the remaining aqueous slurry wasbrought to pH 3 by addition of 150 mL of 1 M H₂SO₄ solution. This wasthen extracted with ethyl acetate (300 mL, 100 mL) and the combinedorganic layers were washed with H₂O (2×) and brine. The organic portionwas dried over Na₂SO₄ and concentrated to give tert-butyl2-(2-hydroxyethoxy)ethylcarbamate as a colorless oil (47.1 g).

Part B

A rapidly stirred solution of tert-butyl2-(2-hydroxyethoxy)ethylcarbamate (47.1 g, 0.230 mol) in 1 L ofanhydrous CH₂Cl₂ was cooled to 0° C. under N₂ and treated withtriethylamine (48.0 mL, 0.345 mol). Methanesulfonyl chloride (19.6 mL,0.253 mol) was then added dropwise over 30 min. The reaction mixture wasthen allowed to warm to room temperature and was stirred an additional22 h. The reaction was quenched by addition of 500 mL saturated NaHCO₃solution and the organic layer was separated. The organic phase was thenwashed with H₂O (3×500 mL) and brine. The organic portion was dried overNa₂SO₄ and concentrated to give2-{2-[(tert-butoxycarbonyl)amino]ethoxy}ethyl methanesulfonate as abrown oil (63.5 g).

Part C

A stirred solution of 2-{2-[(tert-butoxycarbonyl)amino]ethoxy}ethylmethanesulfonate (63.5 g, 0.224 mol) in 400 mL of N,N-dimethylformamide(DMF) was treated with NaN₃ (16.1 g, 0.247 mol) and the reaction mixturewas heated to 90° C. under N₂. After 5 h, the solution was cooled toroom temperature and treated with 500 mL of cold H₂O. The reactionmixture was then extracted with Et₂O (3×300 mL). The combined organicextracts were washed with H₂O (4×100 mL) and brine (2×100 mL). Theorganic portion was dried over MgSO₄ and concentrated to give 52.0 g oftert-butyl 2-(2-azidoethoxy)ethylcarbamate as a light brown oil.

Part D

A solution of tert-butyl 2-(2-azidoethoxy)ethylcarbamate (47.0 g, 0.204mol) in MeOH was treated with 4 g of 10% Pd on carbon and shaken underH₂ (3 Kg/cm²) for 24 h. The solution was then filtered through a Celitepad and concentrated to give 35.3 g of crude tert-butyl2-(2-aminoethoxy)ethylcarbamate as a colorless liquid that was usedwithout further purification.

Part E

A stirred solution of 4-chloro-3-nitroquinoline (31.4 g, 0.151 mol) in500 mL of anhydrous CH₂Cl₂, under N₂, was treated with triethylamine (43mL, 0.308 mol) and tert-butyl 2-(2-aminoethoxy)ethylcarbamate (0.151mol). After stirring overnight, the reaction mixture was washed with H₂O(2×300 mL) and brine (300 mL). The organic portion was dried over Na₂SO₄and concentrated to give a bright yellow solid. Recrystallization fromethyl acetate/hexanes gave 43.6 g of tert-butyl2-{2-[(3-nitroquinolin-4-yl)amino]ethoxy}ethylcarbamate as bright yellowcrystals.

Part F

A solution of tert-butyl2-{2-[(3-nitroquinolin-4-yl)amino]ethoxy}ethylcarbamate (7.52 g, 20.0mmol) in toluene was treated with 1.5 g of 5% Pt on carbon and shakenunder H₂ (3 Kg/cm²) for 24 h. The solution was then filtered through aCelite pad and concentrated to give 6.92 g of crude tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethylcarbamate as a yellowsyrup.

Part G

A solution of tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethylcarbamate (3.46 g, 10.0mmol) in 50 mL of toluene was treated with triethylorthovalerate (2.5mL, 14.5 mmol) and the reaction mixture was heated to reflux. A 25 mgportion of pyridinium hydrochloride was then added and refluxing wascontinued for 4 h. The reaction was then concentrated to dryness underreduced pressure. The residue was dissolved in 50 mL of CH₂Cl₂ andwashed with saturated NaHCO₃, H₂O and brine. The organic portion wasdried over Na₂SO₄ and concetrated to give a green oil. The green oil wasdissolved in 50 mL of hot MeOH and treated with activated charcoal. Thehot solution was filtered and concentrated to give 4.12 g of tert-butyl2-[2-(2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate as ayellow oil.

Part H

A solution of tert-butyl2-[2-(2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate (4.12g, 10.0 mmol) in 50 mL of CH₂Cl₂ was treated with 3-chloroperoxybenzoicacid (MCPBA, 77%, 2.5 g, 11.2 mmol). After stirring for 5 h, thereaction mixture was treated with saturated NaHCO₃ solution and thelayers were separated. The organic portion was washed with H₂O and brinethen dried over Na₂SO₄ and concentrated to give 3.68 g of tert-butyl2-[2-(2-butyl-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamateas a light brown foam.

Part I

A solution of tert-butyl2-[2-(2-butyl-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate(3.68 g, 8.60 mmol) in 100 mL of 1,2-dichloroethane was heated to 80° C.and treated with 10 mL of concentrated NH₄OH solution. To the rapidlystirred solution was added solid p-toluenesulfonyl chloride (1.87 g,9.81 mmol) over a 10 min. period. The reaction mixture was then sealedin a pressure vessel and heating was continued for 2 h. The reactionmixture was then cooled and treated with 100 mL of CH₂Cl₂. The reactionmixture was then washed with H₂O, 1% Na₂CO₃ solution (3×) and brine. Theorganic portion was dried over Na₂SO₄ and concentrated to give 3.68 g oftert-butyl2-[2-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamateas a light brown foam.

Part J

Tert-butyl2-[2-(4-amino-2-butyl-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate(3.68 g, 8.60 mmol) was suspended in 20 mL of 2M HCl in EtOH and themixture was heated to reflux with stirring. After 3 h, the reactionmixture was concentrated to give a solid. The solid was triturated withhot EtOH (50 mL) and filtered to give 2.90 g of the product as thehydrochloride salt. The free base was made by dissolving thehydrochloride salt in 50 mL of H₂O and treating with 5 mL ofconcentrated NH₄OH. The aqueous suspension was extracted with CH₂Cl₂(3×50 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to give1-[2-(2-aminoethoxy)ethyl]-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine asa tan powder.

MS 328 (M+H)⁺;

¹H NMR (300 MHz, CDCl₃) δ 7.95 (d, J=8.3 Hz, 1H); 7.83 (d, J=8.4 Hz,1H); 7.50 (m, 1H); 7.30 (m, 1H); 5.41 (s, 2H); 4.69 (t, J=5.6 Hz, 2H);3.93 (t, J=5.6 Hz, 2H); 3.39 (t, J=5.1 Hz, 2H); 2.97 (t, J=7.9 Hz, 2H);2.76 (t, J=5.1 Hz, 2H); 1.89 (m, 2H); 1.52 (m, 2H); 1.26 (br s, 2H);1.01 (t, J=7.3 Hz, 3H).

EXAMPLE 2 1-[2-(2-aminoethoxy)ethyl]-1H-imidazo[4,5-c]quinolin-4-amine

Part A

A solution of tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethylcarbamate (6.92 g, 20.0mmol) in 100 mL of toluene was treated with triethylorthoformate (4.65mL, 28.0 mmol) and the reaction mixture was heated to reflux. A 100 mgportion of pyridinium hydrochloride was then added and refluxing wascontinued for 2 h. The reaction was then concentrated to dryness underreduced pressure. The residue was dissolved in 200 mL of CH₂Cl₂ andwashed with saturated NaHCO₃, H₂O and brine. The organic portion wasdried over Na₂SO₄ and concentrated to give a green oil. The green oilwas dissolved in 200 mL of hot MeOH and treated with 10 g of activatedcharcoal. The hot solution was filtered and concentrated to give 5.25 gof tert-butyl 2-[2-(1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamateas a light yellow syrup.

Part B

A solution of tert-butyl2-[2-(1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate (5.25 g, 14.7mmol) in 200 mL of CH₂Cl₂ was treated with MCPBA (77%, 3.63 g, 16.3mmol). After stirring overnight, the reaction mixture was treated withsaturated NaHCO₃ solution and the layers were separated. The organicportion was washed with H₂O and brine then dried over Na₂SO₄ andconcentrated to give 4.60 g of tert-butyl2-[2-(5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate as alight brown foam.

Part C

A solution of tert-butyl2-[2-(5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate (4.60g, 12.4 mmol) in 150 mL of 1,2-dichloroethane was heated to 80° C. andtreated with 10 mL of concentrated NH₄OH solution. To the rapidlystirred solution was added solid p-toluenesulfonyl chloride (2.71 g,14.2 mmol) over a 10 min period. The reaction mixture was treated withan additional 2 mL of concentrated NH₄OH solution and then sealed in apressure vessel and heating was continued for 3 h. The reaction mixturewas then cooled and treated with 100 mL of CH₂Cl₂. The reaction mixturewas then washed with H₂O, 1% Na₂CO₃ solution (3×) and brine. The organicportion was dried over Na₂SO₄ and concentrated to give 4.56 g oftert-butyl2-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate as alight brown foam.

Part D

Tert-butyl2-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]ethylcarbamate (4.56g, 12.3 mmol) was dissolved in 100 mL of EtOH and treated with 30 mL of2M HCl in EtOH and the mixture was heated to reflux with stirring. After3 h, the reaction mixture was concentrated to give a solid. The solidwas triturated with hot EtOH (100 mL) and filtered to give the productas the hydrochloride salt. The free base was made by dissolving thehydrochloride salt in 50 mL of H₂O and treating with 5 mL ofconcentrated NH₄OH. The aqueous suspension was extracted with CH₂Cl₂(5×50 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated to give 1.35 g of1-[2-(2-aminoethoxy)ethyl]-1H-imidazo[4,5-c]quinolin-4-amine as a tanpowder.

MS 272 (M+H)⁺;

¹H NMR (300 MHz, CDCl₃) δ 7.98 (d, J=8.2 Hz, 1H); 7.88 (s, 1H); 7.84 (d,J=8.4 Hz, 1H); 7.54 (m, 1H); 7.32 (m, 1H); 5.43 (s, 2H); 4.74 (t, J=5.2Hz, 2H); 3.97 (t, J=5.2 Hz, 2H); 3.42 (t, J=5.1 Hz, 2H); 2.78 (t, J=5.1Hz, 2H); 1.10 (br s, 2H).

EXAMPLE 31-[2-(2-aminoethoxy)ethyl]-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-4-amine

Part A

A solution of tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethylcarbamate (10.2 g, 29.5mmol) in 250 mL of anhydrous CH₂Cl₂ was cooled to 0° C. and treated withtriethylamine (4.18 mL, 30.0 mmol). Methoxypropionyl chloride (3.30 mL,30.3 mmol) was then added dropwise over 5 min.

The reaction was then warmed to room temperature and stirring wascontinued for 1 h. The reaction mixture was then concentrated underreduced pressure to give an orange solid. This was dissolved in 250 mLof EtOH and 12.5 mL of triethylamine was added. The mixture was heatedto reflux and stirred under N₂ overnight. The reaction was thenconcentrated to dryness under reduced pressure and treated with 300 mLof Et₂O. The mixture was then filtered and the filtrate was concentratedunder reduced pressure to give a brown solid. The solid was dissolved in200 mL of hot MeOH and treated with activated charcoal. The hot solutionwas filtered and concentrated to give 11.1 g of tert-butyl2-{2-[2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamateas a yellow syrup.

Part B

A solution of tert-butyl2-{2-[2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamate(10.22 g, 24.7 mmol) in 250 mL of CHCl₃ was treated with MCPBA (77%,9.12 g, 40.8 mmol). After stirring 30 min, the reaction mixture waswashed with 1% Na₂CO₃ solution (2×75 mL) and brine. The organic layerwas then dried over Na₂SO₄ and concentrated to give 10.6 g of tert-butyl2-{2-[2-(2-methoxyethyl)-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamateas an orange foam that was used without further purification.

Part C

A solution of tert-butyl2-{2-[2-(2-methoxyethyl)-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamate(10.6 g, 24.6 mmol) in 100 mL of 1,2-dichloroethane was heated to 60° C.and treated with 10 mL of concentrated NH₄OH solution. To the rapidlystirred solution was added solid p-toluenesulfonyl chloride (7.05 g,37.0 mmol) over a 10 min period. The reaction mixture was treated withan additional 1 mL concentrated NH₄OH solution and then sealed in apressure vessel and heating was continued for 2 h. The reaction mixturewas then cooled and treated with 100 mL of CHCl₃. The reaction mixturewas then washed with H₂O, 1% Na₂CO₃ solution (2×) and brine. The organicportion was dried over Na₂SO₄ and concentrated to give 10.6 g oftert-butyl2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamateas a brown foam.

Part D

Tert-butyl2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethylcarbamate(10.6 g, 24.6 mmol) was treated with 75 mL of 2M HCl in EtOH and themixture was heated to reflux with stirring. After 1.5 h, the reactionmixture was cooled and filtered to give a gummy solid. The solid waswashed EtOH and Et₂O and dried under vacuum to give the hydrochloridesalt as a light brown solid. The free base was made by dissolving thehydrochloride salt in 50 mL of H₂O and treating with 10% NaOH solution.The aqueous suspension was then concentrated to dryness and the residuewas treated with CHCl₃. The resulting salts were removed by filtrationand the filtrate was concentrated to give 3.82 g of1-[2-(2-aminoethoxy)ethyl]-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-4-amineas a tan powder.

MS 330 (M+H)⁺;

¹H NMR (300 MHz, DMSO-d₆) δ 8.10 (d, J=8.1 Hz, 1H); 7.66 (d, J=8.2 Hz,1H); 7.40 (m, 1H); 7.25 (m, 1H); 6.88 (br s, 2H); 4.78 (t, J=5.4 Hz,2H); 3.89 (t, J=4.8 Hz, 2H); 3.84 (t, J=6.9 Hz, 2H); 3.54 (t, J=5.4 Hz,2H); 3.31 (s, 3H); 3.23 (t, J=6.6 Hz, 2H); 2.88 (t, J=5.3 Hz, 2H).

EXAMPLE 4N-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)benzamide

1-[2-(2-Aminoethoxy)ethyl]-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-4-amine(750 mg, 2.28 mmol) was dissolved in 35 mL of anhydrous CH₂Cl₂ andcooled to 0° C. under N₂. To the stirred solution were added Et₃N (0.35mL, 2.50 mmol) and benzoyl chloride (260 μL, 2.28 mmol) and the reactionwas allowed to warm to room temperature over 2.5 h. The reaction mixturewas then quenched by addition of saturated NaHCO₃ solution (30 mL) andCH₂Cl₂ (30 mL). The organic layer was separated and washed with H₂O andbrine, dried over Na₂SO₄ and concentrated under reduced pressure to givea tan foam. Mass spectral analysis showed the presence of some bis-amidein addition to the desired product. The tan foam was treated with 1Naqueous HCl solution (50 mL) at 100° C. for 5 h. HPLC analysis showedthat all of the bis-amide had been converted to the desired product. Thereaction was cooled to room temperature and treated with 10% NaOH untilthe pH˜11. The mixture was extracted with CHCl₃ (3×30 mL). The combinedorganic extracts were washed with H₂O and brine, dried over Na₂SO₄ andconcentrated under reduced pressure to give a yellow solid. Purificationby column chromatography (SiO₂, 5-10% MeOH/CHCl₃) gave 100 mg ofN-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)benzamideas a white powder. m.p. 184-187° C.;

MS 434 (M+H)⁺;

¹H NMR (300 MHz, DMSO-d₆) δ 8.40 (m, 1H); 8.06 (d, J=8.3 Hz, 1H);7.76-7.74 (m, 2H); 7.60 (d, J=7.8 Hz, 1H); 7.54-7.37 (m, 4H); 7.19 (t,J=7.3 Hz, 1H); 6.48 (s, 2H); 4.79-4.72 (m, 2H); 3.91-3.84 (m, 2H); 3.78(t, J=6.9 Hz, 2H); 3.48 (t, J=5.5 Hz, 2H); 3.25 (s, 3H); 3.20 (t, J=6.3Hz, 2H);

¹³C (75 MHz, DMSO-d₆) δ 166.7, 152.0, 151.9, 145.2, 134.8, 132.7, 131.4,128.6, 127.4, 126.7, 121.4, 120.5, 115.1, 70.4, 69.4, 69.2, 58.4, 45.5,27.6.

EXAMPLE 51-[2-(2-aminoethoxy)ethyl]-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine

1-[2-(2-Aminoethoxy)ethyl]-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-4-amine(10.0 g, 27.3 mmol) was dissolved in 50 mL of trifluoroacetic acid andtreated with PtO₂ (1.0 g). The reaction mixture was shaken under H₂ (3Kg/cm²). After 4 d, an additional 0.5 g of PtO₂ was added andhydrogenation was continued for an additional 3 d. The reaction was thenfiltered through Celite and concentrated under reduced pressure to givea brown oil. The yellow oil was dissolved in 200 mL of H₂O then madebasic (pH˜11) by addition of 10% NaOH solution. This was then extractedwith CHCl₃ (5×75 mL) and the combined organic layers were dried overNa₂SO₄ and concentrated to give 5.17 g of1-[2-(2-aminoethoxy)ethyl]-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amineas a tan solid.

MS 334 (M+H)⁺;

¹H NMR (300 MHz, CDCl₃) δ 5.19 (s, 2H); 4.49 (t, J=5.4 Hz, 2H); 3.84 (t,J=6.6 Hz, 2H); 3.71 (t, J=5.4 Hz, 2H), 3.36 (t, J=5.2 Hz, 2H); 3.51 (s,3H); 3.15 (t, J=6.6 Hz, 2H); 2.95 (m, 2H); 2.82 (m, 2H); 2.76 (t, J=5.1Hz, 2H); 1.84 (m, 4H), 1.47 (br s, 2H).

EXAMPLE 6N-(2-{2-[4-amino-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)benzamide

1-[2-(2-Aminoethoxy)ethyl]-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine(1.00 g, 3.00 mmol) was dissolved in 30 mL of anhydrous CH₂Cl₂ andcooled to 0° C. under N₂. To the stirred solution were added Et₃N (0.84mL, 6.00 mmol) and benzoyl chloride (348 μL, 3.00 mmol) and the reactionwas allowed to warm to room temperature overnight. The reaction mixturewas then quenched by addition of saturated NaHCO₃ solution (30 mL). Theorganic layer was separated and washed with H₂O and brine, dried overNa₂SO₄ and concentrated under reduced pressure to give a yellow oil. Theoil was dissolved in a minimum amount of hot MeOH and then treated withEt₂O (50 mL) which caused a white percipitate to form. The solid wasisolated by filtration and dried under vacuum to give 476 mg ofN-(2-{2-[4-amino-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)benzamideas a white powder. m.p. 141-143° C.;

MS 438 (M+H)⁺;

¹H NMR (300 MHz, DMSO-d₆) δ 8.36 (t, J=5.4 Hz, 1H); 7.78-7.76 (m, 2H);7.54-7.42 (m, 3H); 5.68 (s, 2H); 4.43 (t, J=5.4 Hz, 2H); 3.75-3.69 (m,4H); 3.48 (t, J=6.0 Hz, 2H); 3.37 (t, J=5.5 Hz, 2H); 3.24 (s, 3H); 3.07(t, J=6.9 Hz, 2H); 2.91 (m, 2H); 2.63 (m, 2H); 1.70 (m, 4H);

¹³C (75 MHz, DMSO-d₆) δ 166.7, 151.3, 149.3, 146.2, 138.5, 134.8, 131.4,128.6, 127.5, 124.9, 105.6, 70.5, 70.5, 69.3, 58.4, 44.6, 32.7, 27.6,23.8, 23.0, 23.0.

Anal. Calcd for C₂₄H₃₁N₅O₃: % C, 65.88; % H, 7.14; % N, 16.01. Found: %C, 65.55; % H, 7.15; % N, 15.87.

EXAMPLE 72-(2-methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-1H-imidazo[4,5-c]quinolin-4-amine

Part A

Sodium hydride (60% oil dispersion, 9.1 g, 228 mmol) was placed in around bottom flask and washed with hexanes (3×) under N₂. The driedsodium hydride was treated with 800 mL of anhydrous THF. A solution oftert-butyl 2-(2-azidoethoxy)ethylcarbamate (41.9 g, 182 mmol) in 200 mLof THF was then added to the stirred sodium hydride solution over 40min. After addition was complete, the reaction was stirred an additional20 min followed by addition of methyl iodide (13.6 mL, 218 mmol). Afterstirring overnight, the reaction was quenched with 300 mL of saturatedNaHCO₃ solution. The reaction mixture was then treated with 200 mL ofH₂O and 1 L of Et₂O. The organic phase was separated and washed with H₂Oand brine. The organic portion was then dried over MgSO₄ andconcentrated under reduced pressure to give 41.9 g of tert-butyl2-(2-azidoethoxy)ethyl(methyl)carbamate as a yellow liquid.

Part B

A solution tert-butyl 2-(2-azidoethoxy)ethyl(methyl)carbamate (41.9 g,170 mmol) in 600 mL of MeOH was treated with 2.5 g of 10% Pd on carbonand shaken under H₂ (3 Kg/cm²) for 24 h. The solution was then filteredthrough a Celite pad and concentrated to give 37.2 g of crude tert-butyl2-(2-aminoethoxy)ethyl(methyl)carbamate as a light yellow liquid.

Part C

A stirred solution of 4-chloro-3-nitroquinoline (32.3 g, 155 mmol) in400 mL of anhydrous CH₂Cl₂, under N₂, was treated with triethylamine(43.1 mL, 310 mmol) and tert-butyl2-(2-aminoethoxy)ethyl(methyl)carbamate (37.2 g, 171 mmol). Afterstirring overnight, the reaction mixture was washed with H₂O (2×300 mL)and brine (300 mL). The organic portion was dried over Na₂SO₄ andconcentrated to give a brown oil. Column chromatography (SiO₂, 33% ethylacetate/hexanes-67% ethyl acetate/hexanes) gave 46.7 g of tert-butylmethyl(2-{2-[(3-nitroquinolin-4-yl)amino]ethoxy}ethyl)carbamate as ayellow solid.

Part D

A solution of tert-butylmethyl(2-{2-[(3-nitroquinolin-4-yl)amino]ethoxy}ethyl)carbamate (6.56 g,16.8 mmol) in 75 mL of toluene was treated with 0.5 g of 5% Pt on carbonand shaken under H₂ (3 Kg/cm²) for 24 h. The solution was then filteredthrough a Celite pad and concentrated to give 6.8 g of crude tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethyl(methyl)carbamate as anorange syrup which was carried on without further purification.

Part E

A solution of tert-butyl2-{2-[(3-aminoquinolin-4-yl)amino]ethoxy}ethyl(methyl)carbamate (6.05 g,16.8 mmol) in 200 mL of anhydrous CH₂Cl₂ was cooled to 0° C. and treatedwith triethylamine (2.40 mL, 17.2 mmol). Methoxypropionyl chloride (1.72mL, 17.2 mmol) was then added dropwise over 5 min. The reaction was thenwarmed to room temperature and stirring was continued for 3 h. Thereaction mixture was then concentrated under reduced pressure to give anorange solid. This was dissolved in 200 mL of EtOH and 7.2 mL oftriethylamine was added. The mixture was heated to reflux and stirredunder N₂ overnight. The reaction was then concentrated to dryness underreduced pressure and treated with 300 mL of Et₂O. The mixture was thenfiltered and the filtrate was concentrated under reduced pressure togive a brown solid. This was dissolved in 300 mL of CH₂Cl₂ and washedwith H₂O and brine. The organic portion was dried over Na₂SO₄ andconcentrated under reduced pressure to give a brown oil. The oil wasdissolved in 100 mL of hot MeOH and treated with activated charcoal. Thehot solution was filtered and concentrated to give 7.20 g of tert-butyl2-{2-[2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamateas a yellow syrup.

Part F

A solution of tert-butyl2-{2-[2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamate(7.20 g, 16.8 mmol) in 200 mL of CH₂Cl₂ was treated with MCPBA (77%,4.32 g, 19.3 mmol). After stirring 6 h, the reaction mixture was treatedwith saturated NaHCO₃ solution and the layers were separated. Theorganic portion was washed with H₂O and brine then dried over Na₂SO₄ andconcentrated to give 7.05 g of tert-butyl2-{2-[2-(2-methoxyethyl)-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamateas a light brown solid.

Part G

A solution of tert-butyl2-{2-[2-(2-methoxyethyl)-5-oxido-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamate(7.05 g, 15.9 mmol) in 100 mL of 1,2-dichloroethane was heated to 80° C.and treated with 5 mL of concentrated NH₄OH solution. To the rapidlystirred solution was added solid p-toluenesulfonyl chloride (3.33 g,17.5 mmol) over a 10 min period. The reaction mixture was treated withan additional 5 mL concentrated NH₄OH solution and then sealed in apressure vessel and heating was continued for 4 h. The reaction mixturewas then cooled and treated with 100 mL of CH₂Cl₂. The reaction mixturewas then washed with H₂O, 1% Na₂CO₃ solution (3×) and brine. The organicportion was dried over Na₂SO₄ and concentrated to give 6.50 g oftert-butyl2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamateas a brown oil.

Part H

Tert-butyl2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl(methyl)carbamate(6.50 g, 14.7 mmol) was dissolved in 100 mL of EtOH and treated with 20mL of 2M HCl in EtOH and the mixture was heated to reflux with stirring.After 6 h, the reaction mixture was cooled and filtered to give a gummysolid. The solid was washed with EtOH and Et₂O and dried under vacuum togive the hydrochloride salt as a light brown powder. The free base wasmade by dissolving the hydrochloride salt in 50 mL of H₂O and treatingwith 5 mL of concentrated NH₄OH. The aqueous suspension was extractedwith CH₂Cl₂ (5×50 mL). The combined organic layers were dried overNa₂SO₄ and concentrated to give 3.93 g of2-(2-methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-1H-imidazo[4,5-c]quinolin-4-amineas a tan powder.

MS 344 (M+H)⁺;

¹H NMR (300 MHz, DMSO-d₆) δ 8.07 (d, J=7.7 Hz, 1H); 7.62 (dd, J=1.0, 8.3Hz, 1H); 7.42 (ddd, J=1.0, 7.1, 8.2 Hz, 1H); 7.22 (ddd, J=1.1, 7.1, 8.2Hz, 1H); 6.49 (s, 2H); 4.75 (t, J=5.1 Hz, 2H); 3.83 (t, J=6.8 Hz, 4H);3.35 (t, J=5.6 Hz, 2H); 3.30 (s, 3H); 3.21 (t, J=6.9 Hz, 2H); 2.45 (t,J=5.6 Hz, 2H); 2.12 (s, 3H).

EXAMPLE 8N-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)-N-methylbenzamide

2-(2-Methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-1H-imidazo[4,5-c]quinolin-4-amine(1.00 g, 2.92 mmol) was dissolved in 30 mL of anhydrous CH₂Cl₂ andcooled to 0° C. under N₂. To the stirred solution were added Et₃N (0.81mL, 5.81 mmol) and benzoyl chloride (340 μL, 2.92 mmol) and the reactionwas allowed to warm to room temperature overnight. The reaction mixturewas then quenched by addition of saturated NaHCO₃ solution (30 mL) andCH₂Cl₂ (30 mL). The organic layer was separated and washed with H₂O andbrine, dried over Na₂SO₄ and concentrated under reduced pressure.Purification by column chromatography (SiO₂, 3% MeOH/CHCl₃ saturatedwith aqueous NH₄OH) gave the product as a colorless foam.Crystallization from PrOAc and hexanes gave 540 mg ofN-(2-{2-[4-amino-2-(2-methoxyethyl)-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)-N-methylbenzamideas a white powder. m.p. 93.5-97.0° C.;

MS 448 (M+H)⁺;

¹H NMR (500 MHz, DMSO-d₆, 60° C.) δ 8.04 (d, J=7.7 Hz, 1H); 7.63 (dd,J=0.9, 8.2 Hz, 1H); 7.42-7.33 (m, 4H); 7.23-7.19 (m, 3H); 6.24 (s, 2H);4.74 (m, 2H); 3.86 (m, 2H); 3.82 (t, J=6.8 Hz, 2H); 3.51 (m, 2H); 3.40(m, 2H); 3.29 (s, 3H); 3.18 (t, J=6.7 Hz, 2H); 2.75 (br s, 3H);

¹³C NMR (125 MHz, DMSO-d₆, 60° C.) δ 152.0, 151.9, 145.3, 137.1, 132.8,131.3,129.4, 128.5, 127.0, 126.9, 126.8, 126.6, 121.4, 120.4, 115.3,70.5, 69.5, 68.8, 58.4, 45.5, 27.8.

Anal. Calcd for C₂₅H₂₉N₅O₃: % C, 67.09; % H, 6.53; % N, 15.65. Found: %C, 67.08; % H, 6.56; % N, 15.58

EXAMPLE 92-(2-methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine

2-(2-Methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-1H-imidazo[4,5-c]quinolin-4-amine(4.22 g, 12.3 mmol) was dissolved in 25 mL of trifluoroacetic acid andtreated with PtO₂ (0.5 g). The reaction mixture was shaken under H₂ (3Kg/cm²). After 4 d, an additional 0.5 g of PtO₂ was added andhydrogenation was continued for an additional 3 d. The reaction was thenfiltered through Celite and concentrated under reduced pressure to givea yellow oil. The yellow oil was dissolved in 50 mL of H₂O and extractedwith 50 mL of CHCl₃. The organic portion was removed and discarded. Theaqueous portion was then made basic (pH_˜12) by addition of 10% NaOHsolution. This was then extracted with CHCl₃ (6×50 mL) and the combinedorganic layers were dried over Na₂SO₄ and concentrated to a brown oil.The brown oil was dissolved in 100 mL of hot MeOH and treated with 1 gof activated charcoal. The hot solution was filtered through Celite andconcentrated to dryness. The resulting gummy solid was concentratedseveral times with Et₂O to give 3.19 g of2-(2-methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amineas an off-white powder.

MS 348 (M+H)⁺;

¹H NMR (300 MHz, CDCl₃) δ 4.84 (s, 2H); 4.48 (t, J=5.7 Hz, 2H); 3.84 (t,J=6.7 Hz, 2H); 3.70 (t, J=5.7 Hz, 2H); 3.46 (t, J=5.1 Hz, 2H); 3.36 (s,3H); 3.14 (t, J=6.7 Hz, 2H); 2.96 (m, 2H); 2.83 (m, 2H); 2.65 (t, J=5.1Hz, 2H); 2.36 (s, 3H); 1.85 (m, 4H).

EXAMPLE 10N-(2-{2-[4-amino-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)-N-methylbenzamide

2-(2-Methoxyethyl)-1-{2-[2-(methylamino)ethoxy]ethyl}-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-4-amine(750 mg, 2.16 mmol) was dissolved in 20 mL of anhydrous CH₂Cl₂ andcooled to 0° C. under N₂. To the stirred solution were added Et₃N (0.60mL, 4.32 mmol) and benzoyl chloride (250 μL, 2.16 mmol) and the reactionwas allowed to warm to room temperature overnight. The reaction mixturewas then quenched by addition of saturated NaHCO₃ solution (30 mL) andCH₂Cl₂ (30 mL). The organic layer was separated and washed with H₂O (3×)and brine, dried over Na₂SO₄ and concentrated under reduced pressure.Purification by column chromatography (SiO₂, 3% MeOH/CHCl₃ saturatedwith aqueous NH₄OH) gave the product as a colorless foam. The foam wasconcentrated from iso-propyl alcohol to give an syrup which solidifiedupon standing. The solid was dried under vacuum to give the 408 mg ofN-(2-{2-[4-amino-2-(2-methoxyethyl)-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolin-1-yl]ethoxy}ethyl)-N-methylbenzamideas an off-white powder. m.p 83.0-87.0° C.;

MS 452 (M+H)⁺;

¹H NMR (500 MHz, DMSO-d₆, 60° C.) δ 7.37 (m, 3H); 7.23 (m, 2H); 5.46 (s,2H); 4.43 (m, 2H); 3.76 (t, J=6.8 Hz, 2H); 3.68 (m, 2H); 3.50 (m, 2H);3.42 (m, 2H); 3.27 (s, 3H); 3.05 (t, J=6.4 Hz, 2H); 2.92 (m, 2H); 2.80(s, 3H); 2.65 (m, 2H); 1.74 (m, 4H);

¹³C NMR (125 MHz, DMSO-d₆, 60° C.) δ 150.5, 148.5, 145.8, 137.9, 136.4,128.7, 127.8, 126.3, 124.5, 105.1, 70.1, 69.8, 68.0, 57.7, 44.0, 32.1,27.1, 23.2, 22.4, 22.4

Anal. Calcd for C₂₅H₃₃N₅O₃0.30C₃H₈O: % C, 66.24; % H, 7.60; % N, 14.91.Found: % C, 65.86; % H, 7.81; % N, 15.10.

EXAMPLE 111-{1-[(2-piperidin-4-ylethoxy)methyl]propyl}-1H-imidazo[4,5-c]quinolin-4-amine

Part A

Using the general method of Part A of Example 1,4-piperidineethanol (10g, 77.4 mmol) was reacted with di-tert-butyl dicarbonate (17.7 g, 81.3mmol) to provide 13.1 g of tert-butyl4-(2-hydroxyethyl)piperidine-1-carboxylate as a clear oil.

Part B

Iodine (7.97 g) was added in three portions to a solution of imidazole(3.89 g, 57.1 mmol) and triphenylphosphine (14.98 g, 57.1 mmol) indichloromethane (350 mL). After 5 minutes a solution of the materialfrom Part A in dichloromethane (70 mL) was added. The reaction mixturewas stirred at ambient temperature overnight. More iodine (7.97 g) wasadded and the reaction was stirred at ambient temperature for 1 hr. Thereaction mixture was washed with saturated sodium thiosulfate (2×) andbrine, dried over sodium sulfate, filtered and then concentrated underreduced pressure to provide an oily residue. The residue was purified bycolumn chromatography (silica gel eluting with 20% ethyl acetate inhexanes) to provide 15.52 g of tert-butyl4-(2-iodoethyl)piperidine-1-carboxylate as a pale yellow oil.

Part C

Under a nitrogen atmosphere,2-(1H-imidazo[4,5-c]quinolin-1-yl)butan-1-ol (6.5 g, 26.9 mmol) wasadded in three portions to a suspension of sodium hydride (1.4 g of 60%,35.0 mmol) in anhydrous N,N-dimethylformamide. The reaction mixture wasallowed to stir for 45 minutes by which time gas evolution had ceased.Tert-butyl 4-(2-iodoethyl)piperidine-1-carboxylate (10.05 g, 29.6 mmol)was added dropwise over a period of 15 minutes. The reaction mixture wasallowed to stir at ambient temperature for 2.5 hrs; then it was heatedto 100° C. and stirred overnight. Analysis by HPLC showed that thereaction was about 35% complete. Saturated ammonium chloride solutionwas added, the resulting mixture was allowed to stir for 20 minutes andthen it was extracted with ethyl acetate (2×). The ethyl acetateextracts were washed with water (2×) and then with brine, combined,dried over sodium sulfate, filtered and then concentrated under reducedpressure to provide a brown oil. The oil was purified by columnchromatography (silica gel eluting sequentially with 30% ethyl acetatein hexanes, 50% ethyl acetate in hexanes, and ethyl acetate) to provide2.2 g of tert-butyl4-{2-[2-(1H-imidazo[4,5-c]quinolin-1-yl)butoxy]ethyl}piperidine-1-carboxylate.

Part D

Using the general method of Example 1 Part H, the material from Part Cwas oxidized to provide tert-butyl4-{2-[2-(5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)butoxy]ethyl}piperidine-1-carboxylateas an oil.

Part E

Ammonium hydroxide solution (20 mL) was added to a solution of thematerial from Part D in dichloromethane (20 mL). A solution of tosylchloride (0.99 g, 5.2 mmol) in dichloromethane (10 mL) was added over aperiod of 5 minutes. The resulting biphasic reaction mixture was allowedto stir overnight. The reaction mixture was diluted with chloroform andsaturated sodium bicarbonate solution. The layers were separated. Theorganic layer was dried over sodium sulfate, filtered and thenconcentrated under reduced pressure to provide a brown glass. Thismaterial was purified by column chromatography (silica gel eluting firstwith 50% ethyl acetate in hexanes and then with ethyl acetate) toprovide 1.0 g of tert-butyl4-{2-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)butoxy]ethyl}piperidine-1-carboxylateas pale yellow glassy foam.

Part F

Under a nitrogen atmosphere, tert-butyl4-{2-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)butoxy]ethyl}piperidine-1-carboxylate(1.00 g, 2.1 mmol) and 2N ethanolic hydrochloric acid (10 ml, 20 mmol)were combined and the solution was stirred at ambient temperature for 14hours. The solvent was removed in vacuo and the resulting tan solid wasdissolved in water. Saturated aqueous sodium carbonate was added untilthe pH reached 10. After extraction with dichloromethane (3×), theorganic fractions were combined, washed with brine, dried (Na₂SO₄),filtered, and the majority of the solvent was removed in vacuo. Hexanewas added to form a precipitate. Vacuum filtration yielded 0.5 g of1-{1-[(2-piperidin-4-ylethoxy)methyl]propyl}-1H-imidazo[4,5-c]quinolin-4-amineas a tan powder.

¹H-NMR (300 MHz, DMSO-d₆) δ 8.34 (bs, 1H), 8.19 (d, J=8.49 Hz, 1H), 7.61(dd, J=8.31, 1.13 Hz, 1H), 7.45-7.39 (m, 1H), 7.25-7.19 (m, 1H), 6.55(s, 2H), 5.25-5.15 (m, 1H), 4.00-3.80 (m, 2H), 3.5-3.3 (m, 2H), 2.8-2.64(m, 2H), 2.22-2.11 (m, 2H), 2.09-1.99 (m, 2H), 1.8-1.63 (bs, 1H),1.37-1.0 (m, 5H), 0.95-0.7 (m, 5H);

¹³ C-NMR (75 MHz, DMSO-d₆): δ 152.8, 145.8, 140.6, 133.0, 127.8, 127.0,126.9, 121.3, 121.0,115.5, 71.8, 68.1, 58.4,46.1, 36.3, 33.1, 32.7,24.5, 9.9;

MS (CI) m/e 368.2459 (368.2450 calcd for C₂₁H₃₀N₅O).

EXAMPLE 125-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]-N-methyl-N-phenylpentanamide

Using the general method of Parts C and D of Example 11,2-(1H-imidazo[4,5-c]quinolin-1-yl)ethanol (0.63 g, 2.9 mmol) and5-bromo-N-methyl-N-phenylpentanamide (1.3 g, 4.8 mmol) were combined toprovide 0.24 g of5-[2-(5-oxido-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]-N-methyl-N-phenylpentanamideas a colorless oil. The resulting N-oxide product was dissolved indichloromethane and trichloroacetyl isocyanate (0.11 ml) was addeddropwise. The reaction was stirred at room temperature for 2 hours andthen the solvent was removed under vacuum. The resulting oil wasdissolved in methanol and sodium methoxide (0.2 ml, 25% by weight inmethanol) was slowly added. The reaction was maintained overnight andthen concentrated under vacuum. Purification by flash columnchromatography (silica gel, 9:1 ethyl acetate\methanol) provided 24 mgof5-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]-N-methyl-N-phenylpentanamideas a white solid.

¹H NMR (300 MHz, CDCl₃) δ 7.94 (d, J=8.1 Hz, 1H), 7.83 (m, 2H), 7.52(dt, J=7.7, 1.3 Hz, 1H), 7.41-7.28 (m, 4H), 7.12 (d, J=7.8 Hz, 2H), 5.55(broad s, 2H), 4.65 (t, J=5.3 Hz, 2H), 3.85 (t, J=5.3 Hz, 2H), 3.31 (t,J=6.3 Hz, 2H), 3.24 (s, 3H), 2.02 (m, 2H), 1.56 (m, 2H), 1.40 (m, 2H);

IR (KBr) 3429, 3104, 2946, 2877, 1646, 1595, 1584, 1532, 1496, 1482,1398, 1360, 1254, 1121, 749, 705 cm⁻¹;

MS (EI) m/e 417.2160 (417.2165 calcd for C₂₄H₂₇N₅O₂).

EXAMPLE 135-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]-N-butyl-N-phenylpentanamide

2-(1H-Imidazo[4,5-c]quinolin-1-yl)ethanol and5-bromo-N-butyl-N-phenylpentanamide were combined and treated accordingto the general procedure described in Example 12. Purification by flashcolumn chromatography (silica gel, 98:2 ethyl acetate\methanol) provided5-[2-(4-amino-1H-imidazo[4,5-c]quinolin-1-yl)ethoxy]-N-butyl-N-phenylpentamideas a colorless oil.

¹H NMR (300 MHz, CDCl₃) δ 7.93 (d, J=8.0 Hz, 1H), 7.87-7.85 (m, 2H),7.54 (dt, J=7.7, 1.1 Hz, 1H), 7.41-7.29 (m, 4H), 7.10 (d, J=7.2 Hz, 2H),6.20 (broad s, 2H), 4.66 (t, J=5.2 Hz, 2H), 3.85 (t, J=5.2 Hz, 2H), 3.66(t, J=7.5 Hz, 2H), 3.31 (t, J=6.2 Hz, 2H), 1.96 (t, J=7.2 Hz, 2H),1.56-1.25 (m, 8H), 0.88 (t, J=7.2 Hz, 3H);

MS (EI) m/e 459.2631 (459.2634 calcd for C₂₇H₃₃N₅O₂).

EXAMPLE 14 Methyl[2-(4-Amino-1H-imidazo[4,5-c]quinolin-1-yl)butoxy]acetate

2-(4-Amino-1H-imidazo[4,5-c]quinolin-1-yl)butan-1-ol (25 mg, 0.0975mmol) was placed in a 2 dram (7.4 mL) vial. Sodium hydride (5 mg of a60% dispersion in mineral oil, 0.117 mmol) and N,N-dimethylformamide (1mL) were added. The vial was placed on a sonicator for 15 minutes atambient temperature to allow the alkoxide to form. Methyl bromoacetate(11 μL, 0.117 mmol) was added. The reaction was sonicated at ambienttemperature for 1.5 hours. The reaction mixture was analyzed by LC/MS toconfirm the formation of the desired product. The reaction mixture waspurified by semi-preparative HPLC using Method A Mass Measurement (Da.):Theoretical mass=328.1535, Measured mass=328.1534.

EXAMPLES 15-34

The compounds in the table below were prepared according to thesynthetic method of step (7) of Reaction Scheme II above using thefollowing general method.

The acid chloride (84 μmol) was added to a test tube containing asolution of1-[2-(2-aminoethoxy)ethyl]-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine (25mg, 77 μmol) in dichloromethane (5 mL). The test tube was capped andthen placed on a shaker at ambient temperature for 20 hr. The solventwas removed by vacuum centrifugation. The residue was purified bysemi-preparative HPLC using Method B described above. The table belowshows the structure of the free base and the observed ac curate mass(M+H).

Example Accurate Number Structure of Free Base Mass (obs.) 15

396.2404 16

412.2717 17

424.2717 18

432.2407 19

438.2748 20

446.2560 21

450.2318 22

452.2116 23

457.2369 24

457.2333 25

460.2693 26

462.2529 27

462.2502 28

467.1952 29

472.2708 30

476.2659 31

482.2568 32

500.2246 33

500.2252 34

516.2217

EXAMPLES 35-51

The compounds in the table below were prepared according to thesynthetic method of step (7) of Reaction Scheme II above using thefollowing general method.

The acid chloride (1.1 eq.) was added to a test tube containing asolution of1-{1-[(2-piperidin-4-ylethoxy)methyl]propyl}-1H-imidazo[4,5-c]quinolin-4-amine(25 mg) in dichloromethane (5 mL). The test tube was capped and thenplaced on a shaker at ambient temperature for 20 hr. The solvent wasremoved by vacuum centrifugation. The residue was purified bysemi-preparative HPLC using Method B described above. The table belowshows the structure of the free base and the observed accurate mass(M+H).

Example Accurate Mass Number Structure of Free Base (obs.) 35

436.2683 36

452.3014 37

464.3045 38

472.2717 39

486.2878 40

490.2592 41

492.2419 42

497.2682 43

500.3022 44

502.2824 45

507.2281 46

473.2686 47

512.3038 48

516.2965 49

522.2836 50

540.2534 51

556.2521

EXAMPLES 52-66

The compounds in the table below were prepared according to thesynthetic method of step (7) of Reaction Scheme II above using thefollowing general method.

1-[2-(2-Aminoethoxy)ethyl]-1H-imidazo[4,5-c]quinolin-4-amine (20 mg) and1-methyl-2-pyrrolidinone (5 mL) were combined in a test tube andsonicated with heating to provide a solution. The acid chloride (1.1eq.) was added. The test tube was capped and then placed on a shaker atambient temperature for 20 hr. The solvent was removed by vacuumcentrifugation. The residue was purified by semi-preparative HPLC usingMethod B described above. The table below shows the structure of thefree base and the observed accurate mass (M+H).

Example Accurate Mass Number Structure of Free Base (obs.) 52

356.2093 53

376.1783 54

382.2260 55

390.1949 56

396.1503 57

401.1707 58

404.2105 59

406.1855 60

406.1861 61

411.1320 62

420.2047 63

426.1912 64

444.1628 65

444.1655 66

460.1608

Cytokine Induction in Human Cells

An in vitro human blood cell system is used to assess cytokineinduction. Activity is based on the measurement of interferon and tumornecrosis factor (α) (IFN and TNF, respectively) secreted into culturemedia as described by Testerman et. al. In “Cytokine Induction by theImmunomodulators Imiquimod and S-27609”, Journal of Leukocyte Biology,58, 365-372 (September, 1995).

Blood Cell Preparation for Culture

Whole blood from healthy human donors is collected by venipuncture intoEDTA vacutainer tubes. Peripheral blood mononuclear cells (PBMCs) areseparated from whole blood by density gradient centrifugation usingHistopaque®-1077. The PBMCs are washed twice with Hank's Balanced SaltsSolution and then are suspended at 3-4×10⁶ cells/mL in RPMI complete.The PBMC suspension is added to 48 well flat bottom sterile tissueculture plates (Costar, Cambridge, Mass. or Becton Dickinson Labware,Lincoln Park, N.J.) containing an equal volume of RPMI complete mediacontaining test compound.

Compound Preparation

The compounds are solubilized in dimethyl sulfoxide (DMSO). The DMSOconcentration should not exceed a final concentration of 1% for additionto the culture wells

Incubation

The solution of test compound is added to the first well containing RPMIcomplete and serial dilutions are made in the wells. The PBMC suspensionis then added to the wells in an equal volume, bringing the testcompound concentrations to the desired range. The final concentration ofPBMC suspension is 1.5-2×10⁶ cells/mL. The plates are covered withsterile plastic lids, mixed gently and then incubated for 18 to 24 hoursat 37° C. in a 5% carbon dioxide atmosphere.

Separation

Following incubation the plates are centrifuged for 5-10 minutes at 1000rpm (˜200× g) at 4° C. The cell-free culture supernatant is removed witha sterile polypropylene pipet and transferred to sterile polypropylenetubes. Samples are maintained at −30 to −70° C. until analysis. Thesamples are analyzed for interferon (α) and for tumor necrosis factor(α) by ELISA.

Interferon (α) and Tumor Necrosis Factor (α) Analysis by ELISA

Interferon (α) concentration is determined by ELISA using a HumanMulti-Species kit from PBL Biomedical Laboratories, New Brunswick, N.J.Results are expressed in pg/mL.

Tumor necrosis factor (α) (TNF) concentration is determined using ELISAkits available from Genzyme, Cambridge, Mass.; R&D Systems, Minneapolis,Minn.; or Pharmingen, San Diego, Calif. Results are expressed in pg/mL.

The table below lists the lowest concentration found to induceinterferon and the lowest concentration found to induce tumor necrosisfactor for each compound. A “*” indicates that no induction was seen atany of the tested concentrations; generally the highest concentrationtested was 10 or 30 μM.

Cytokine Induction in Human Cells Example Lowest Effective Concentration(μM) Number Interferon Tumor Necrosis Factor 12 3.33 * 13 10 * 14 0.37 *15 0.1  1 16 0.1  1 17 1  1 18 1 10 19 1 10 20 0.1 10 21 1 10 22 0.1 1023 1 10 24 1 10 25 1 10 26 1 10 27 1 10 28 1 10 29 1 10 30 1 10 31 * 1032 * 10 33 * 10 34 * 10 35 0.1  1 36 1  1 37 1  1 38 1 10 39 1 10 40 1 1 41 0.1  1 42 1  1 43 1 10 44 1  1 45 0.1  1 46 0.1  1 47 1 * 48 0.110 49 1 10 50 1  1 51 10 10 52 10 10 53 10 10 54 10 10 55 1 10 56 1 1057 10 * 58 10 10 59 10 10 60 10 10 61 10 10 62 1 10 63 * 10 64 10 *65 * * 66 * *

What is claimed is:
 1. A compound of the formula (I):

wherein: X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-; R₁ is selectedfrom the group consisting of: —R₄—CR₃—Z—R₆-alkyl; —R₄—CR₃—Z—R₆-alkenyl;—R₄—CR₃—Z—R₆-aryl; —R₄—CR₃—Z—R₆-heteroaryl; —R₄—CR₃—Z—R₆-heterocyclyl;R₄—CR₃—Z—H; —R₄—NR₇—CR₃—R₆-alkyl; —R₄—NR₇CR₃—R₆-alkenyl;—R₄—NR₇—CR₃—R₆-aryl; —R₄—NR₇—CR₃—R₆-heteroaryl;—R₄—NR₇—CR₃—R₆-heterocyclyl; and —R₄—NR₇—CR₃—R₈; each Z is independently—NR₅—, —O—, or —S—; R₂ is selected from the group consisting of:-hydrogen; -alkyl; -alkenyl; -aryl; -heteroaryl; -heterocyclyl;-alkyl-Y-alkyl; -alkyl-Y-alkenyl; -alkyl-Y-aryl; and -alkyl or alkenylsubstituted by one or more substituents selected from the groupconsisting of: —OH; -halogen; —N(R₅)₂; —CO—N(R₅)₂; —CO—C₁₋₁₀alkyl;—CO—O—C₁₋₁₀alkyl; —N₃; -aryl -heteroaryl; -heterocyclyl; —CO-aryl; and—CO-heteroaryl; each R₃ is ═O or ═S; each R₄ is independently alkyl oralkenyl, which may be interrupted by one or more —O— groups; each R₅ isindependently H or C₁₋₁₀alkyl; R₆ is a bond, alkyl, or alkenyl, whichmay be interrupted by one or more —O— groups; R₇ is H, C₁₋₁₀ alkyl, orarylalkyl; or when R₄ is alkyl and R₇ is C₁₋₁₀ alkyl, R₄ and R₇ can jointogether to form a piperidine ring; R₈ is H or C₁₋₁₀ alkyl; each Y isindependently —O— or —S(O)₀₋₂—; n is 0 to 4; and each R present isindependently selected from the group consisting of C₁₋₁₀ alkyl, C₁₋₁₀alkoxy, hydroxy, halogen and trifluoromethyl; or a pharmaceuticallyacceptable salt thereof.
 2. A compound or salt of claim 1 wherein theheteroaryl is selected from the group consisting of 2-pyridyl,3-pyridyl, 4-pyridyl, 2-thiazolyl, and 4-pyrazolyl.
 3. A compound orsalt of claim 1 wherein X is —CH(alkyl)(alkyl)-wherein the alkyl groupscan be the same or different.
 4. A compound or salt of claim 1 wherein Xis —CH₂—CH₂—.
 5. A compound or salt of claim 1 wherein X is—CH(C₂H₅)(CH₂)—.
 6. A compound or salt of claim 1 wherein R₂ is H.
 7. Acompound or salt of claim 1 wherein R₂ is alkyl.
 8. A compound or saltof claim 1 wherein R₂ is -alkyl-O-alkyl.
 9. A compound of the formula(II)

wherein: X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-; R₁ is selectedfrom the group consisting of: —R₄—CR₃—Z—R₆-alkyl; —R₄—CR₃—Z—R₆-alkenyl;—R₄—CR₃—Z—R₆-aryl; —R₄—CR₃—Z—R₆-heteroaryl; —R₄—CR₃—Z—R₆-heterocyclyl;—R₄—CR₃—Z—H; —R₄—NR₇—CR₃—R₆-alkyl; —R₄—NR₇—R₆-alkenyl;—R₄—NR₇—CR₃—R₆-aryl; —R₄—NR₇—CR₃—R₆—heteroaryl;—R₄—NR₇—CR₃—R₆-heterocyclyl; and —R₄—NR₇—CR₃—R₈; each Z is independently—NR₅—, —O—, or —S—; R₂ is selected from the group consisting of:-hydrogen; -alkyl; -alkenyl; -aryl; -heteroaryl; -heterocyclyl;-alkyl-Y-alkyl; -alkyl-Y-alkenyl; -alkyl-Y-aryl; and -alkyl or alkenylsubstituted by one or more substituents selected from the groupconsisting of: —OH; -halogen; —N(R₅)₂; —CO—N(R₅)₂; —CO—C₁₋₁₀ alkyl;—CO—O—C₁₋₁₀ alkyl; —N₃; aryl; -heteroaryl; -heterocyclyl; —CO-aryl; and—CO-heteroaryl; each R₃ is ═O or ═S; each R₄ is independently alkyl oralkenyl, which may be interrupted by one or more —O— groups; each ₅ isindependently H or C₁₋₁₀ alkyl; ₆ is a bond, alkyl, or alkenyl, whichmay be interrupted by one or more —O— groups; R₇ is H, C₁₋₁₀ alkyl, orarylalkyl; or when R₄ is alkyl and R₇ is C₁₋₁₀ alkyl, R₄ and R₇ can jointogether to form a piperidine ring; R₈ is H or C₁₋₁₀ alkyl; each Y isindependently —O— or —S(O)₀₋₂—; n is 0 to 4; and each R present isindependently selected from the group consisting of C₁₋₁₀ alkyl, C₁₋₁₀alkoxy, hydroxy, halogen, and trifluoromethyl; or a pharmaceuticallyacceptable salt thereof.
 10. A compound or salt of claim 9 wherein R₂ isH or alkyl.
 11. A compound or salt of claim 9 wherein R₂ is-alkyl-O-alkyl.
 12. A pharmaccutical composition comprising atherapeutically effective amount of a compound or salt of claim 1 and apharmaceutically acceptable carrier.
 13. A method of inducing cytokinebiosynthesis in an animal comprising administering a therapeuticallyeffective amount of a compound or salt of claim 1 to the animal.
 14. Themethod of claim 13 wherein the cytokine is IFN-α.
 15. A method oftreating a viral disease in an animal in need thereof comprisingadministering to the animal a therapeutically effective amount of acompound or salt of claim 1 that induces cytokine biosynthesis.
 16. Acompound of the formula (III):

wherein: X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl; R₁ is selectedfrom the group consisting of: —R₄—CR₃—Z—R₆—alkyl; —R₄—CR₃—Z—R₆—alkenyl;—R₄—CR₃—Z—R₆-aryl; —R₄—CR₃—Z—R₆-heteroaryl; —R₄—CR₃—Z—R₆-heterocyclyl;—R₄—CR₃—Z—H; —R₄—NR₇—CR₃—R₆-alkyl; —R₄—NR₇—CR₃—R₆-alkenyl;—R₄—NR₇—CR₃—R₆-aryl; —R₄—NR₇—CR₃—R₆-heteroaryl;—R₄—NR₇—CR₃—R₆-heterocyclyl; and —R₄—NR₇—CR₃—R₈; each Z is independently—NR₅—, —O—, or —S—; R₂ is selected from the group consisting of:-hydrogen; -alkyl; -alkenyl; -aryl; -heteroaryl; -heterocyclyl;-alkyl-Y-alkyl; -alkyl-Y-alkenyl; -alkyl-Y-aryl; and -alkyl or alkenylsubstituted by one or more substituents selected from the groupconsisting of: —OH; -halogen; —N(R₅)₂; —CO—N(R₅)₂; —CO—C₁₋₁₀ alkyl;—CO—O—C₁₋₁₀ alkyl; —N₃; aryl; -heteroaryl; -heterocyclyl; —CO-aryl; and—CO-heteroaryl; each R₃ is ═O or ═S; each R₄ is independently alkyl oralkenyl, which may be interrupted by one or more —O— groups; each R₅ isindependently H or C₁₋₁₀ alkyl; R₆ is a bond, or is alkyl, or alkenyl,which may be interrupted by one or more —O— groups; R₇ is H, C₁₋₁₀alkyl, or arylalkyl; or when R₄ is alkyl and R7 is C₁₋₁₀ alkyl, R₄ andR₇ can join together to form a piperidine ring; R₈ is H or C₁₋₁₀ alkyl;each Y is independently —O— or —S(O)₀₋₂—; n is 0 to 4; and each Rpresent is independently selected from the group consisting of C₁₋₁₀alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen and trifluoromethyl; or apharmaceutically acceptable salt thereof.
 17. A compound of the formula(IV):

wherein X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-; R₁ is selectedfrom the group consisting of: —R₄—CR₃—Q—R₆-alkyl; —R₄—CR₃—Q—R₆-alkenyl;—R₄—CR₃—Q—R₆-aryl; —R₄—CR₃—Q—R₆-heteroaryl; —R₄—CR₃—Q—R₆-heterocyclyl;—R₄—CR₃—Q—H; —R₄—NR₅—CR₃—R₆-alkyl; —R₄—NR₅—CR₃—R₆-alkenyl;—R₄—NR₇—CR₃—R₆-aryl; —R₄—NR₇—CR₃—R₆-heteroaryl;—R₄—NR₇—CR₃—R₆-heterocyclyl; and —R₄—NR₇—CR₃—R₈; each Q is independently—NR₅— or —O—; each R₃ is ═O or ═S; each R₄ is independently alkyl oralkenyl, which may be interrupted by one or more —O— groups; each R₅ isindependently H or C₁₋₂₀ alkyl; R₆ is a bond, alkyl, or alkenyl, whichmay be interrupted by one or more —O— groups; R₇ is H, C₁₋₁₀ alkyl, orarylalkyl; or when R₄ is alkyl and R₇ is C₁₋₁₀ alkyl, R₄ and R₇ can jointogether to form a piperidine ring; R₈ is H or C₁₋₁₀ alkyl; n is 0 to 4;and each R present is independently selected from the group consistingof C₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, halogen and trifluoromethyl; or apharmaceutically acceptable salt thereof.
 18. A pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundor salt of claim 9 and a pharmaceutically acceptable carrier.
 19. Amethod of inducing cytokine biosynthesis in an animal comprisingadministering a therapeutically effective amount of a compound or saltof claim 9 to the animal.
 20. The method of claim 19 wherein thecytokine is IFN-α.
 21. A method of treating a viral disease in an animalin need thereof comprising administering to the animal a therapeuticallyeffective amount of a compound or salt of claim 9 that induces cytokinebiosynthesis.
 22. A compound of the formula (V):

wherein: X is —CHR₅—, —CHR₅-alkyl-, or —CHR₅-alkenyl-; R₂ is selectedfrom the group consisting of: -hydrogen; -alkyl; -alkenyl; -aryl;-heteroaryl; -heterocyclyl; -alkyl-Y-alkyl; -alkyl-Y-alkenyl;-alkyl-Y-aryl; and -alkyl or alkenyl substituted by one or moresubstituents selected from the group consisting of: —OH; -halogen;—N(R₅)₂; —CO—N(R₅)₂; —CO—C₁₋₁₀ alkyl; —CO—O—C_(1.10) alkyl; —N₃; -aryl;-heteroaryl; -heterocyclyl; —CO-aryl; and —CO-heteroaryl; each R₄ isindependently alkyl or alkenyl, which may be interrupted by one or more—O— groups; R₇ is H, C₁₋₁₀ alkyl, or arylalkyl; or when R₄ is alkyl andR₇ is C₁₋₁₀ alkyl, R₄ and R₇ can join together to form a piperidinering; each Y is independently —O— or —S(O)₀₋₂—; n is 0 to 4; and each Rpresent is independently selected from the group consisting of C₁₋₁₀alkyl, C₁₋₁₀ alkoxy, hydroxy, halogen and trifluoromethyl; or apharmaceutically acceptable salt thereof.
 23. The method according toclaim 13 wherein the animal has a viral disease.
 24. The methodaccording to claim 13 wherein the animal has a neoplastic disease. 25.The method according to claim 19 wherein the animal has a viral disease.26. The method according to claim 19 wherein the animal has a neoplasticdisease.